Clustered regularly interspaced short palindromic repeats (CRISPR) ke mokhoa oa ho hlophisa liphatsa tsa lefutso tsa moloko oa boraro o fetotseng lefats'e ka liphetho tsa oona tse phahameng. O sebelisitsoe ho phekola mafu a fapaneng a baeloji le tšoaetso. Libaktheria tse fapaneng le li-prokaryotes tse ling (tse kang archaea) le tsona li na le litsamaiso tsa CRISPR/Cas9 ho itšireletsa khahlanong le li-phages. Ho tlalehiloe hore maano a thehiloeng ho CRISPR/Cas9 a ka thibela kholo le tsoelo-pele ea mofetše oa matsoele o nang le triple negative (TNBC) ka ho shebana le liphatsa tsa lefutso tse ka fetoloang tsa ho hanyetsa, transcription, le taolo ea epigenetic. Mekhoa ena ea phekolo e ka thusa ho rarolla mathata a thata joalo ka ho hanyetsa lithethefatsi ho bonoang esita le ho TNBC. Hona joale, mekhoa e fapaneng e sebelisoa ho isa CRISPR/Cas9 liseleng tse shebaneng, joalo ka mekhoa ea 'mele (microinjection, electroporation le hydrodynamic), vaerase (vaerase e amanang le adeno le lentivirus) le e seng ea vaerase (liposomes le lipid nanoparticles). Leha mehlala e fapaneng e ntlafalitsoe ho ithuta lisosa tsa limolek'hule tsa TNBC, khaello ea mekhoa e bonolo le e shebaneng le eona ea ho fana ka lisebelisoa tsa ho hlophisa liphatsa tsa lefutso ka har'a 'mele e fokotsa ts'ebeliso ea tsona ea bongaka. Ka bokhutšoanyane, tlhahlobo ena e hlahloba ka botlalo tsoelo-pele, liphephetso, mefokolo le menyetla ea kalafo ea CRISPR/Cas9 bakeng sa TNBC ho latela bopaki bo seng bo ntse bo le teng. Re boetse re totobatsa kamoo motsoako oa bohlale ba maiketsetso le ho ithuta ha mochini ho ka ntlafatsang maano a CRISPR/Cas9 kalafong ea TNBC.
Kankere e khetholloa ka karohano e sa laoleheng ea lisele, tšitiso ea libaka tsa tlhahlobo ea potoloho ea sele, le liphetoho liphatseng tsa lefutso tse hatellang hlahala (TSGs) (Matthews et al., 2022). Har'a mefuta e fapaneng ea kankere, kankere ea matsoele ke mofuta o tloaelehileng haholo oa kankere ho basali 'me o na le sekhahla se phahameng sa lefu lefatšeng ka bophara (Waks le Winer, 2019). Kankere ea matsoele ke lefu le fapaneng le nang le litšobotsi tse fapaneng, tse kang likarolo tsa histological le tsa baeloji, tlhahiso ea kliniki le boitšoaro, le karabelo kalafong (Weigelt et al., 2010). Ho arola kankere ea matsoele ho fana ka temohisiso e nepahetseng bakeng sa tlhahlobo ea kankere ea matsoele le tebello ea hlahala. Tšebeliso ea li-biomarker tse tloaelehileng le likarolo tsa kliniki e bile tekanyetso e ka sehloohong ea ho arola kankere ea matsoele (Tsang le Tse, 2019). Tebello le karabelo kalafong ea kankere ea matsoele li susumetsoa ke lintlha tse ngata, ho kenyeletsoa boteng ba estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2/neu), le histological grade, mofuta oa hlahala, le sehlopha. Metastasis ea boholo le li-lymph node (Al-Tubaiti, 2020). Mefuta e mehlano ea limolek'hule tsa kankere ea matsoele e fumanoe, ho kenyeletsoa luminal A, luminal B, HER2-enriched, basal-like, le claudin-low (Prat et al., 2015). TNBC ke mofuta oa limolek'hule oa kankere o sa hlahiseng ER, PR, le HER2 kaofela (Yin et al., 2020). Litšobotsi tsena tsa pathological li amahanngoa le TNBC, e tiisang tsoelo-pele ea eona e potlakileng le tlhaho e mabifi ho feta mofuta ofe kapa ofe oa kankere ea matsoele (Feng et al., 2018). Ho phaella moo, Peru et al (2000) ba sebelisitse theknoloji ea microarray ho arola bocha kankere ea matsoele le ho khetholla mefuta e mehlano ea limolek'hule tsa kankere ea matsoele (Cadenas, 2012). Kankere ea matsoele e kang ea basal ke mofuta oa kankere ea matsoele o nang le phenotype e mpe ka makhetlo a mararo 'me o amahanngoa le tsoelo-pele e potlakileng. Ho lokela ho hlokomeloa hore kankere tsohle tsa matsoele tse kang tsa basal hangata li fumanoa hampe e le TNBC, empa ke 77% feela e leng TNBC; Ka lehlakoreng le leng, 71-91% ea TNBC e tšoana le ea basal, e leng se bontšang hore mefuta ena e 'meli ea kankere ea matsoele ea kopana 'me e emela lihlopha tse fapaneng (Wang D.-Y. et al., 2019). Sena se etsa hore ho be le tlhoko ea ho hlalosa ho se tšoane ha TNBC ho hlakisa tebello le ho khetholla likarabo tse ka bang teng kalafong ea hona joale le ea nakong e tlang. Ho phaella moo, TNBC e ikarabella bakeng sa 15-20% ea linyeoe tsohle tsa kankere ea matsoele 'me e atile haholo ho basali ba ka tlase ho lilemo tse 50. Liphetoho tsa BRCA1 kapa BRCA2 li tlalehiloe hoo e ka bang 20% ea linyeoe tsa TNBC (Xie et al., 2017; Tzikas et al., 2017). , 2020). Lithuto li boetse li bontšitse hore TNBC e na le tikoloho e ikhethang ea 'mele e kenyeletsang maemo a holimo a lintho tse bakang kholo ea methapo ea mali, li-macrophage tse amanang le hlahala (li-TAM), li-lymphocyte tse kenang ka har'a hlahala (li-TIL), le limolek'hule tse ling tse amehang kholong le ho falla ha hlahala. Ka hona, ho utloisisa tikoloho e nyenyane ea TNBC ho bohlokoa bakeng sa tebello ea eona le kalafo (Fan le He, 2022).
Ho lekola tebello ea TNBC le ho netefatsa kalafo e sebetsang, tlhahlobo e nepahetseng, haholo-holo e thehiloeng ho immunohistochemistry (IHC) ho lemoha ER, PR le HER2, le mammography ho lemoha li-neoplasm tsa matsoele, ke habohlokoa. Leha ho le joalo, mammography e ke ke ea bontša ka ho lekaneng litšobotsi tsa ka hare ho hlahala tse kang necrosis le fibrosis (Deepak Singh et al., 2021). Kaha tebello e mpe ea tebello le tlhahlobo li thibela lingaka ho fana ka meriana e nepahetseng (Chaudhary, 2020), ho sebelisoa maano a fapaneng ho ntlafatsa tlhokomelo ea bakuli ba nang le TNBC. Hajoale, meriana e 'meli, doxorubicin le cyclophosphamide, e sebelisoa ho bakuli ba nang le TNBC 'me e bontšitse liphetho tse ntle. Ho phaella moo, ho sebelisoa meriana e meng ea platinum, joalo ka carboplatin le cisplatin (Sikov et al., 2015). Ho phaella moo, li-inhibitors tsa PARP tse kang Olaparib, Velaparib, le PF-01367338 li sebelisitsoe e le meriana e ka bang teng ea chemotherapy bakeng sa kalafo ea TNBC (Ishino et al., 2018). Kaha ho tlalehiloe hore litsela tsa ho supa tsa Wnt/b-Catenin, NOTCH, le Hedgehog li ameha ho hlaheng le ho tsoeleng pele ha TNBC, ho lebisa meriana litseleng tsena e ka ba leano la bohlokoa (Aysola et al., 2013). Leha opereishene, phekolo ea mahlaseli le chemotherapy e ntse e le motheo oa kalafo bakeng sa TNBC kajeno, ho entsoe tsoelo-pele e kholo ho nts'etsapele mekhoa e mecha ea phekolo ho kenyeletsoa phekolo e reretsoeng, immunotherapy, lisebelisoa tse fapaneng tsa ho hlophisa liphatsa tsa lefutso tse amanang le CRISPR tse kang Cas9n, dCas9., CRISPR/Cas12, prime editing, le phekolo ea liphatsa tsa lefutso e reretsoeng CRISPR/Cas9. Mona re tla tsepamisa maikutlo ho lisebelisoa tse fapaneng tsa ho hlophisa liphatsa tsa lefutso tse amanang le CRISPR tse sebelisoang kalafong ea TNBC. Har'a tsona, CRISPR/Cas9 e fumane tlhokomelo e pharalletseng.
Cas9n, e tsejoang hape e le Cas9 nickase, ke mofuta o entsoeng ka liphatsa tsa lefutso oa protheine ea Cas9 e nkiloeng tsamaisong ea ho hlophisa genome ea CRISPR/Cas9 (Gupta et al., 2019). Ka sebopeho sa eona sa pele, protheine ea Cas9 e na le libaka tse peli tsa nuclease: RuvC le HNH, mosebetsi oa eona o ka sehloohong ke ho arohana ha likhoele ka bobeli tsa DNA. Leha ho le joalo, ho Cas9n, e 'ngoe ea libaka tsa nuclease, HNH, e fetoloa ka liphatsa tsa lefutso 'me e fetoha e sa sebetseng. Ka hona, sebaka sa HNH sa Cas9n se lula se sa sebetse. Ke sebaka sa RuvC feela se lulang se sebetsa, se lumellang Cas9n ho khaola kapa ho etsa likheo holim'a likhoele tse le 'ngoe tsa DNA (Trevino le Zhang, 2014). Ha e bapisoa le Cas9, Cas9n e ka fokotsa liphello tse seng tsa sepheo ka katleho le ho ntlafatsa ka nepo mekhoa ea ho lokisa lisele. Cas9n e ka sebelisoa ho rarolla mathata a fapaneng, joalo ka ho etsa likheo libakeng tse itseng ho DNA e nang le likhoele tse peli. Bakeng sa morero ona, limolek'hule tse peli tsa Cas9n li ile tsa kopanngoa 'me tsa sebelisoa hammoho (Yee, 2016). Mixed lineage kinase 3 (MLK3) ke protheine kinase e sebetsang ka mitogen e sebetsang e le molaoli oa bohlokoa nakong ea metastasis ea TNBC (Cronan et al., 2012).
MLK3 e ka kenya tshebetsong ditsela tse ngata tsa ho bontsha tse lebisang ho metastasis ya TNBC. Mohlala, tsela ya c-Jun N-terminal kinase (JNK) e laola motility ya sele le ho senyeha ha matrix ya kantle ho sele, mme MLK3 e kenya tshebetsong tsela ya JNK, ka hona e ntlafatsa motility ya sele le ho fana ka thepa e hlaselang (Rattanasinchai le Gallo, 2016). MLK3 e boetse e laola EMT. Ho etsa sena, e kenya tshebetsong dintho tse latelang tsa ho ngola tse kang Snail, Slug le Twist. Dintlha tsena di thibela tlhahiso ya matshwao a epithelial mme di kgothaletsa tlhahiso ya matshwao a mesenchymal, e leng se lebisang ho fumaneng phenotype e metastatic (Casalino et al., 2023). MLK3 e bonwe e bapala karolo ho ECM ho ntlafatseng le ho kentsweng tshebetsong ha diprotheine tse kang matrix metalloproteinases (MMPs) tse senyang ECM. Sena se nolofalletsa tlhaselo ya sele ya hlahala le ho hasana libakeng tse hole (Katari et al., 2019). Kahoo, dithuto tse fetileng di bontshitse hore MLK3 e bapala karolo ya bohlokwa ho TNBC. Rattanasinchai le Gallo ba sebelisitse mehlala ea TNBC ho ithuta karolo ea MLK3 'me ba fumane hore e khothalletsa nts'etsopele ea mofetše ka litsela tse itseng tsa lipontšo. Ba sebelisitse CRISPR/Cas9n ho hlophisa MLK3 'me ba bone phokotso e kholo ho metastasis ea TNBC (Rattanasinchai le Gallo, 2016).
dCas9, eo hangata e bitsoang Cas9 e sa sebetseng, ke mofuta o fetotsoeng oa protheine ea Cas9. Ho fapana le Cas9 e sebetsang, dCas9 ha e na ts'ebetso ea endonuclease, e leng se etsang hore e se khone ho baka likheo tsa DNA tse peli. Ka hona, dCas9 e ka sebelisoa ho shebana hantle le libaka tse itseng tsa genome ntle le liphetoho kapa liphetoho ho tatellano ea DNA (Wang et al., 2016). Ho dCas9, liprotheine tse peli tsa endonuclease, RuvC le HNH, lia emisoa ka ho thibela masala a bohlokoa a amino acid. (Richter et al., 2016). Ho sa tsotellehe khaello ea eona ea ts'ebetso ea ho arola DNA, dCas9 e bapala karolo tse 'maloa tsa bohlokoa lipatlisisong tsa liphatsa tsa lefutso le theknoloji ea baeloji. Mohlala, e lumella ho bona libaka tse itseng tsa genome, e nolofatsa taolo ea mongolo, 'me e khothalletsa liphetoho tsa epigenetic (Brocken et al., 2018).
Ntho ea ho ngola ea ZEB1 (lebokose la E le tlamang la monwana oa zinki 1) e bapala karolo e ikhethang le ea bohlokoa ho buisaneng le phetoho ea epithelial-mesenchymal (EMT), ts'ebetso ea sele e etsahalang nakong ea nts'etsopele ea lesea le sa tsoa emoloa, tokiso ea lisele, le tsoelo-pele ea kankere. E kenyelletsa phetoho ea lisele tsa epithelial ho ba lisele tsa mesenchymal, e leng se fellang ka liphetoho mofuteng oa sele, motility, invasion, jj. (Wu et al., 2020). ZEB1 e thibela matšoao a mangata a epithelial, joalo ka E-cadherin le Occludin, tse ikarabellang bakeng sa ho boloka ho khomarela ha sele le polarity ea lisele tsa epithelial ho TNBC (Moreno-Bueno et al., 2008). Ho phaella moo, ZEB1 e kenya tšebetsong matšoao a itseng a kang N-cadherin, vimentin le fibronectin (Konradi et al., 2014) mme e boetse e laola liphatsa tsa lefutso tse amehang ho tsosolosoeng ha cytoskeletal joalo ka Rho GTPases le matrix metalloproteinases (MMPs) (Huang). et al., 2014). 2022), ho phaella moo, e boetse e ama ditsela tse fapaneng tsa ho bontsha tse kang ho fetola kgolo ya ntho-β (TGF-β), Wnt signaling, jj., ka hona e kgothaletsa tlhaselo ya hlahala le ho ata ha metastasis ho TNBC (Chen et al., 2016). ). Diphuputso tse ngata le bopaki ba saense di bontsha hore ZEB1 e na le bokgoni bo boholo e le moemedi wa bohlokwa bakeng sa ho lemoha le ho kenella kalafong ya TNBC. Phuputsong ya morao tjena, Waryah et al. ba sebedisitse mohlala wa TNBC mme ba fihlella kgatello e felletseng ya ZEB1 ke dCas9. Kahoo, ba bone ho hlaka ho hoholo haholo le thibelo e batlang e feletse ya ZEB1 maemong a mmele (Waryah et al., 2023).
CRISPR/Cas12 ke sesebelisoa sa ho hlophisa liphatsa tsa lefutso se bitsoang CRISPR/Cpf1. Se nkiloe tsamaisong ea CRISPR/Cas, moo lentsoe Cas12 le bolelang protheine e amanang le CRISPR 12 (Bharathkumar et al., 2022), e tšoanang ka ho feletseng le Cas9. Phapang feela ke hore e na le protheine ea Cas12. Ha e bapisoa le Cas9, Cas12 e na le mesebetsi e ikhethang joalo ka ho hlahisa lipheletso tse khomarelang nakong ea ts'ebetso ea ho hlophisa liphatsa tsa lefutso, athe Cas9 e hlahisa lipheletso tse sa hlakang (Wang et al., 2021). Thepa ena ea Cas12 e thusa theknolojing ea eona e ikhethang ea ho laola DNA. Joaloka protheine e khonang ho lemoha le ho khaola DNA e reretsoeng ka nepo, e na le mekhoa e mengata e ikhethang, e etsang hore e be sesebelisoa se sebetsang bakeng sa lits'ebetso tse fapaneng, ho kenyeletsoa le ho hlophisa liphatsa tsa lefutso (Pickar-Oliver le Gersbach, 2019).
Ho tšoana le mefuta e meng ea CRISPR, CRISPR/Cas12 e boetse e khona ho tlosa kapa ho kenya tšebetsong liphatsa tsa lefutso ho TNBC, e shebane le liphatsa tsa lefutso tse bapalang karolo ea bohlokoa ho pathogenesis ea eona kapa karabelo ea kalafo (Yang le Zhang, 2023). Ho phethahatsa ts'ebetso ena, RNA e tataisang (gRNA) e ile ea hlahisoa e lebisang Cas12 ho gene e reretsoeng. Hang ha Cas12 e tlama ho sepheo sa eona, e hlahisa likheo tse peli tsa DNA 'me e kenya tšebetsong mekhoa ea ho lokisa DNA. Nakong ea ts'ebetso ea tokiso, li-nucleotide tse fosahetseng li ka kenyelletsoa, e leng se lebisang liphetohong tsa liphatsa tsa lefutso le tahlehelo ea ts'ebetso (Zhang et al., 2021a). Ho phaella moo, mofuta o ntlafalitsoeng oa enzyme ea Cas12 (e bitsoang dCas12) le oona o ile oa sebelisoa ho kenya tšebetsong liphatsa tsa lefutso. Ka ho e kopanya le activator ea transcription, hoa khoneha ho susumetsa liphatsa tse itseng tsa lefutso, ka hona ho li kenya tšebetsong. Theknoloji ena e na le bokhoni bo boholo ba ho khothaletsa ts'ebetso ea liphatsa tsa lefutso tse hatellang hlahala (Sultan et al., 2022).
Ho hlophisa ka sehloohong ke theknoloji e sa tsoa ntlafatsoa le e phahameng ka ho fetisisa ea ho hlophisa liphatsa tsa lefutso. E khona ho fetola DNA ea sebopuoa ka nepo haholo (Chen le Liu, 2023). Ho hlophisa ka sehloohong ho finyelloa ka ho kopanya likarolo tse peli tse kholo: enzyme e fetotsoeng ea CRISPR/Cas9 le reverse transcriptase. Karolo ea enzyme ea CRISPR/Cas9 ke ho shebana le libaka tse nepahetseng ka har'a genome ka khetho, ha reverse transcriptase e thusa ho fetola DNA ka botlalo sebakeng seo e reretsoeng ho sona (Hassan et al., 2021). Mokhoeng oa ho hlophisa ka sehloohong, mohato oa pele o kenyelletsa ho thehoa ha RNA ea tataiso ea ho hlophisa ka pele (pegRNA) (Standage-Beier et al., 2021). E na le tatellano ea sepheo le template ea RNA e tsamaellanang le sebaka sa ho hlophisa se lakatsehang ho DNA e reretsoeng. Ebe pegRNA e kenngoa liseleng tse reretsoeng hammoho le primer editing nuclease (PE2). PE2 ke protheine ea fusion e nang le enzyme ea Cas9, reverse transcriptase, le adaptara ea ho hlophisa ka pele (Martín-Alonso et al., 2021). Ka hare ho sele, di-complex tsa pegRNA le PE2 di batla DNA e ikgethang eo di batlang ho e fetola. Enzyme ya Cas9 e seha DNA mme e etsa template e nang le di-strains tse le nngwe (Choi et al., 2022). Reverse transcriptase e sebedisa template ena ho lokisa DNA bakeng sa ho hlophiswa. Hammoho le ho kopitsa DNA, ditaelo tsa ho hlophisa template ya RNA le tsona di a kenyeletswa. Qetellong, khoele ya DNA e sa tswa thehwa e sebediswa e le template ho lokisa lekhalo ho DNA, e leng se fellang ka tatellano ya DNA e fetotsweng e nang le phetoho e lakatsehang (Ochoa-Sanchez et al., 2021). Mekgwa ya ho hlophisa pele e ka lebisa diphetohong tse atileng diphatseng tsa lefutso. E ka lebisa diphetohong tsa ntlha, ho kenngwa, ho hlakolwa, esita le ho nkelwa sebaka ha diphatsa tsa lefutso (Anzalone et al., 2019; Chen le Liu, 2023). Ho hlophisa pele ho fana ka melemo e mmalwa ho feta mahlale a fetileng a ho hlophisa genome. Tsena di kenyeletsa ho nepahala ho eketsehileng, ditlamorao tse fokotsehileng tse sa rerelwang, le bokgoni ba ho hlophisa DNA ntle le ho itshetleha ka di-strands tse pedi tsa DNA (Anzalone et al., 2020).
Theknoloji ea CRISPR/Cas9 e qalile ho ntshetswa pele ho sireletsa libaktheria phetisong ea plasmid le tšoaetsong ea phage 'me hamorao e ile ea sebelisoa hape e le sesebelisoa se sebetsang sa ho shebana le DNA se thehiloeng ho RNA bakeng sa ho hlophisa genome (Jiang le Doudna, 2017). Ho phaella moo, tsamaiso ea CRISPR/Cas9 e tlalehiloe e le teng ho 50% le 87% ea li-genome tsa baktheria le tsa khale, ka ho latellana (Ishino et al., 2018). CRISPR/Cas9 ke sesebelisoa se ka bang teng sa ho hlakola, ho kenya le ho lokisa tatellano efe kapa efe e sa tloaelehang ea liphatsa tsa lefutso ho sebelisoa mekhoa ea in vivo le in vitro (Sabit et al., 2021). Ho feta moo, CRISPR/Cas9 e bontšitsoe e le karolo ea tsamaiso ea 'mele ea ho itšireletsa mafung e ikamahanyang le maemo ka lebaka la ho ikhetha ha eona bakeng sa liphatsa tsa lefutso tse reriloeng (Chen le Zhang, 2018).
CRISPR/Cas9 e na le Cas9 le RNA e le 'ngoe e tataisang (sgRNA). Cas9 ke endonuclease e entsoeng ka likarolo tse ngata tsa protheine. Ho phaella moo, Cas9 e na le thepa e ikhethang ea sebopeho le sebopeho (Pacesa et al., 2022) kaha e na le li-lobe tse peli: ho lemoha (REC) le nuclease (NUC). Lobe ea REC e arotsoe hape ka libaka tse tharo: REC1, REC2, le li-helice tsa borokho (Cromwell et al., 2018). NUC e na le li-lobe tse tharo, e leng RuvC (RuvC I, RuvC II, RuvC III), HNH le sebaka sa tšebelisano sa protospacer accumulated motif (PAM) (Setšoantšo sa 1) (Song et al., 2016). sgRNA e na le likarolo tse peli, ho kenyeletsoa CRISPR RNA (crRNA) le transcoding small RNA (tracer RNA) (Setšoantšo sa 1). SgRNA e hokahanya protheine ea Cas9 le li-connexin ho theha motsoako o sebetsang, o tsejoang hape e le motsoako oa effector (Richter et al., 2012). crRNA ke para ea motheo ea nucleotide ea 18-20 e bapalang karolo ea bohlokoa ho lemoheng tatellano ea DNA e reretsoeng. Ho phaella moo, crRNA e tsamaisana le DNA e reretsoeng, 'me tracrRNA e sebetsa e le scaffold bakeng sa nuclease ea Cas9 ho tlama DNA e reretsoeng (Manghwar et al., 2019). Ka lehlakoreng le leng, tatellano ea PAM e na le li-nucleotide tse 3, tse tiisang, tse hlalosang le ho buisana ka ho tlama ha motsoako oa effector ho DNA. Li-subunit tsa Cas9 complex RuvC le HNH li na le ts'ebetso ea catalytic (Richter et al., 2012; Asmamaw le Zawdie, 2021). Sebaka sa nuclease sa HNH sa subunit ea Cas9 se arola khoele ea DNA e amanang le crRNA. Sebaka sa RuvC nuclease se kgaola dithapo tse ding tsa DNA mme se hlahisa dikgeo tsa dithapo tse pedi (DSBs), ka mora moo mekgwa e mmedi e fapaneng ya ho lokisa dikgeo tsa DNA e kenngwa tshebetsong (Jiang le Doudna, 2017) (Setshwantsho sa 1).
Setšoantšo sa 1. Kakaretso ea CRISPR/Cas9 (A). Likarolo tsa sistimi ea CRISPR/Cas9: (i). Cas9 endonuclease e ikarabella bakeng sa ho khaola tatellano ea DNA e reriloeng, (ii) RNA e le 'ngoe ea tataiso (sg) e bakoang ke ho kopanngoa ha crRNA le tra-crRNA chimeras. (tse peli). Cas9 e kenyelletsa likarolo tse 'maloa tse kang Rec I, Rec II, NUC lobe (HNH le Ruv C ke likaroloana) le sebaka sa tšebelisano sa PAM (C) se nang le mesebetsi e tsamaellanang. Motsoako oa protheine oa CRISPR/Cas9 o arola tatellano ea DNA ka mefuta e seng e tlatsanang le e tlatsanang (D). CRISPR/Cas9 e fetola genome ka mekhahlelo e meraro: ho lemoha, ho khaola le ho lokisa. Sg-RNA e entsoeng e tataisa Cas9 mme e lemoha tatellano e lakatsehang ka crRNA ea karolo e tlatsanang. Cas9 e lemoha tatellano ea PAM ho 5′-NGG-3′ mme e qhibilihisa DNA, e etsa motsoako oa DNA-RNA le ho kenya tšebetsong karohano. Sebaka sa HNH sa Cas9 se kgaola khoele e tlatsanang, mme sebaka sa RuvC se kgaola khoele e sa tlatsanang. CRISPR/Cas9 e lokisa dsDNA ka ho e roba ka ditsela tse pedi: ho kopanya pheletso e sa tlatseletsaneng (NHEJ) le ho lokisa ho tsamaisanang (HDR). NHEJ e lokisa DNA e nang le dithapo tse pedi ha ho se na DNA e tswang kantle ho naha ka tshebetso ya enzymatic, mokgwa o sekametseng ho diphoso o ka kenyang kapa wa tlosa ditatelano tsa DNA tse sa reroang. HDR e ikgethile haholo mme e hloka ditempele tsa DNA tse tshwanang.
Litsela tsa tokiso tse tsamaisoang ke CRISPR/Cas9 li kenyelletsa mekhoa ea ho kopanya qetello e seng ea homologous (NHEJ) le ho lokisa e tataisoang ke homology (HDR). NHEJ ke tsela e sekametseng ho etsa liphoso hobane e kenyelletsa ho kenngoa kapa ho hlakoloa 'me ha e hloke paterone efe kapa efe nakong ea mokhoa oa tokiso. E sebelisa li-nucleotide tse sa reroang ho hlahisa liprotheine tse tloaelehileng (Abbasi et al., 2021). Sistimi ena ea tokiso e na le li-complex tse 'ne tse kang KU complex, cross-complementing protein complex type 4 (XRCC-4), enzyme ea ho sebetsana le DNA end, le protein kinase DNA-PKcs (Abbasi et al., 2021). Protheine e rarahaneng ea KU e na le li-subunit tse peli, Ku 70 le Ku 80, 'me e bapala karolo ea bohlokoa mokhoeng oa NHEJ, kaha mokhoa oa ho lokisa o qalisoa ke ho tlama li-subunit tse peli (Ku 70 le Ku 80) ho lipheletso tse bohlasoa kapa tse batlang li le bohlasoa tsa DNA e reriloeng (Abbasi et al., 2021), e sebetsang e le sethala sa ho bokella lintho tse ling tse amanang le NHEJ sebakeng sa kotsi (Yang et al., 2020). XRCC-4 le ligase ea DNA li entsoe ka li-amino acid tse 334 le 911, ka ho latellana, 'me motsoako oa XRCC4-DNA ligase IV o susumetsa ho hokahana ha lipheletso tsa DNA (Chatterjee et al., 2015). Enzyme ea ho sebetsana le DNA end, e tsejoang hape e le polynucleotide kinase 3′-phosphate, ke enzyme ea ho sebetsana le DNA end. E ka tlosa sehlopha sa 3′P ho DNA le ho fosforisa sehlopha sa 5′OH nakong ea tokiso ea DSB. E boetse e kenyelletsa ho lokisoa ha likheo tsa khoele e le 'ngoe (SSBs) ho sebelisoa tsela ea tokiso ea SSB (Chatterjee et al., 2015). Protein kinase DNA-PKcs ke protheine kinase e itšetlehileng ka DNA e nang le karoloana e nyane ea catalytic ea PIKKs (phosphatidylinositol 3-kinase-related kinases) le lelapa la ataxia telangiectasia mutated (ATM), hammoho le ATM le Rad3-related ATRs. likheo tsa khoele (DSBs) le likheo tsa khoele e le 'ngoe (Yue et al., 2020; Peng et al., 2016).
HDR ke mokhoa o nepahetseng le o loketseng oa ho lokisa hobane tlhahisoleseling e kopitsoa ho sebelisoa mokhoa o sa senyehang oa duplex ea DNA e tšoanang, leha e hloka boteng ba li-chromatids tsa khaitseli. Sena se etsahala nakong ea mohato oa S/G2 oa potoloho ea sele ea mammalian. HDR e hlaha haholo-holo mefuteng ea tomoso, empa NHEJ e bohlokoa ho liphoofolo tse anyesang (Burma et al., 2006; Abbasi et al., 2021). Mokhoa o felletseng oa ho lokisa o bontšitsoe ho Setšoantšo sa 1.
Kaha TNBC e bakoa ke ho se tloaelehe ha liphatsa tsa lefutso le tsa epigenetic, tokiso ea ho se tloaelehe ha liphatsa tsa lefutso tse kotsi/epigenomic ho sebelisoa CRISPR/Cas9 e ka ba mokhoa o utloahalang oa phekolo (Chen et al., 2019). Ho phaella moo, lintlha tse ling tsa ho ngola tse amehang taolong ea ho ngola e khethehileng ea sele li ka bontša litšobotsi tse ikhethang tsa lisele tsa mofetše, e leng se bontšang hore taolo ea ho ngola e ka ba mokhoa o motle oa kalafo ea mofetše (Drost et al., 2017). Ho sebelisa litšobotsi tsena tsa limolek'hule tsa lihlahala, tse kang liphoso tsa liphatsa tsa lefutso, tsa epigenetic le tsa ho ngola, bakeng sa nts'etsopele ea lithethefatsi ho ka ntlafatsa liphetho tsa bongaka le ho fokotsa litšenyehelo tsa tlhahlobo. CRISPR ke sesebelisoa se ka bang teng sa ho hlophisa liphatsa tsa lefutso se ke keng sa lemoha le ho khetholla lipheo tsa genomic tse bakang mofetše feela, empa hape se ka sebelisoa ho hlophisa, ho hatella le ho fetola li-oncogene ka epigenetic liseleng tsa batho (Tafole ea 1) (Ahmed et al., 2021). Ho entsoe liteko tse 'maloa tsa CRISPR ho batla liphatsa tsa lefutso tse amanang le li-tumor suppressors, oncogenes, le ho hanyetsa lithethefatsi. Tšebeliso ea CRISPR/Cas9 ho hlophisa li-oncogene tse fapaneng tsa TNBC e bontšitsoe ho Setšoantšo sa 2.
Setšoantšo sa 2. Ho hlophisoa ha liphatsa tsa lefutso tsa li-oncogene tse fapaneng tsa TNBC ho sebelisoa CRISPR/Cas9 ho fellang ka ho fokotseha ha kholo ea hlahala le ho ata ha eona. Li-oncogene tsena li kenyelletsa CDK7, NAT1, UBR5, YTHDF2, ITGA9, CXCR4 le CXCR7, Crypto1, ROR1 le ST8SIA, tse amehang ho hlaheng le ho ata ha TNBC.
Ho falla ha lisele tsa mofetše, tlhaselo, le phetoho ea epithelial-mesenchymal (EMT) li amahanngoa le ITGA9. Lithuto tse fetileng li bontšitse hore ITGA9 ke sebapali sa bohlokoa tseleng ea Notch 'me e bapala karolo e khahlisang ho metastasis ea rhabdomyosarcoma (Molist et al., 2020). Ho phaella moo, ITGA9 e fumanoe e amana haufi-ufi le tebello ea mokuli mefuteng e 'maloa ea hlahala, ho kenyeletsoa kankere ea matsoele (Wang Z. et al., 2019). Ho phaella moo, tlhahlobo ea bioinformatics ea ITGA9 e bontšitse hore tlhahiso ea eona ho TNBC e ne e le holimo haholo ho feta mefuteng e meng ea kankere ea matsoele. Maemo a phahameng a ITGA9 a amahanngoa le metastasis ea hlahala le ho khutlela morao ho bakuli ba TNBC. Ho tlosoa ha ITGA9 ke CRISPR/Cas9 ho bakile thepa e kang ea sele ea mofetše (CSC), angiogenesis ea hlahala, kholo ea hlahala, le ho fokotseha ha metastasis ka ho khothalletsa ho senyeha ha β-catenin ho TNBC (Wang Z. et al., 2019).
Crypto-1 ke setho sa lelapa la TGF-β mme e bohlokwa bakeng sa ho qala ha embryonic, tlhokomelo ya disele tsa stem, le metastasis ya kankere (Ishii et al., 2021). E tsejwang hape e le Tdgf-1, ke protheine ya ho bontsha e nang le oncogenic GPI e amehang taolong ya ho thehwa ha mothapo wa pele, mesoderm le endoderm, hammoho le ho thehwa ha asymmetry ya letshehadi/le letona ntshetsopeleng ya ditho tsa mmele nakong ya embryonic (Zhang et al., 2021b). Ho feta moo, Cripto-1 e bontshitswe e ameha phetohong ya epithelial-mesenchymal (EMT) e le letshwao la sele ya stem (Zhang et al., 2021b). EMT ha e bohlokwa feela bakeng sa mekgwa e fapaneng e kang ntshetsopele ya embryonic, fibrosis le phodiso ya maqeba, empa hape le bakeng sa ho kenella ha kankere le metastasis. Ho feta moo, Cripto-1 e bontshitswe e sebelisana le di-receptor tse nne tsa Notch mme e ntlafatsa kgolo ya tsona ya kamora phetolelo (Brandstadter le Maillard, 2019). Tsela ea ho supa Notch e tsebahala e ameha tlhokomelong ea lisele tsa kankere ea matsoele tsa batho. Lithuto li bontšitse hore ho tlosoa ha Crypto-1 ho bakoang ke CRISPR/Cas9 ho thibela kholo ea kankere le ho ata ha eona. Ka hona, Crypto-1 e ka ba sepheo sa bohlokoa sa kalafo bakeng sa TNBC (Castro et al., 2015).
Protheine ea XCL12 le receptor ea eona ea chemokine CXC (CXCR4 le CXCR7) li bapala karolo e fapaneng ho atiseng ha lisele tsa mofetše, kholong, ho falleng le ho hlaseloa (Wu et al., 2015). Li-chemoreceptor tsena li 'nile tsa amahanngoa le nts'etsopele ea TNBC ka litsela tse ngata tsa lipontšo ka bobeli mehlala ea in vivo le in vitro (Wu et al., 2015). Ho phaella moo, ho kengoa tšebetsong ha CXCR4 le CXCR7 ho amahanngoa le ho ba kotsing e kholo ea ho ata ha lisele le tebello e mpe ea lefu ho TNBC (Karn et al., 2022). Ka hona, ho tlosoa ha CXCR4 le CXCR7 ho ka fetoha liphatsa tsa lefutso tse sebetsang hantle bakeng sa kalafo ea mofetše oa matsoele, ho kenyeletsoa le TNBC. Phuputso ea Yang et al. Yang et al. (2019) e sebelisitse CRISPR/Cas9 ho kopanya liphatsa tsa lefutso tsa CXCR4 le CXCR7 'me ea fumana hore ho ata ha lisele tsa mofetše, kholo, ho falla le ho hlaseloa ha tsona ho ne ho thibetsoe haholo (Yang et al., 2019).
Maemo a eketsehileng a miR-3662, e leng oncogene ea TNBC, a bonoe liseleng tsa kankere ea matsoele (Yi et al., 2022). Ho kokobetsoa ha miR-3662 ho bontšitsoe ho thibela kholo ea hlahala ea kankere ea matsoele le ho ata ha eona ka har'a 'mele le ka har'a 'mele (Agarwal le Gupta, 2021). HBP-1 ke thibelo e matla ea matšoao a Wnt/-catenin 'me ho ka etsahala hore e ikarabelle bakeng sa kholo ea lisele tsa TNBC e tsamaisoang ke miR-3662. Haufinyane, Yi et al. ba fumane hore axis ea miR-3662-HBP1 e laola tsela ea matšoao a Wnt/-catenin liseleng tsa TNBC (Yi et al., 2022). Ka lebaka la polelo ea eona e ikhethang ea hlahala, miR-3662 e ka ba sepheo se ka bang teng sa kalafo bakeng sa TNBC. Kahoo, ho kokobetsoa ha miR-3662 ho tsamaisoang ke CRISPR/Cas9 e ka ba mokhoa o motle oa ho nts'etsapele meriana e mecha bakeng sa kalafo ea TNBC.
UBR5 ke nucleophosphoprotein ea 300 kDa e 'nileng ea khetholloa e le molaoli oa bohlokoa oa ho hlaha ha tumorigenesis, metastasis, le karabelo ea' mele ea ho itšireletsa mafung ho mefuta e fapaneng ea kankere (Shearer et al., 2015; Fu et al., 2023). Ho tlalehiloe hore UBR5 e eketsehile haholo lisampoleng tsa TNBC 'me e hlasimolla ts'ebetso ea ERα ka ho baka keketseho ka ts'ebetso ea eona ea ubiquitin ligase (Bolt et al., 2015). Lithuto tsohle tsa tatellano ea exome tsa lisampole tsa mantlha tsa TNBC le tsona li bontšitse keketseho ea tlhahiso ea UBR5, e leng se bontšang karolo ea eona nts'etsopele ea TNBC. Ho phaella moo, ho tlosoa ha UBR5 ho susumetsoang ke CRISPR/Cas9 ho bontšitse thibelo e kholo ea metastasis ea TNBC le kholo mehlaleng ea liteko tsa litoeba. Ho feta moo, ho kenyelletsoa ha UBR5 mohlaleng oa litoeba tsa mofuta oa hlaha ho khutlisitse ts'ebetso ea eona e felletseng, athe phello ena ha ea ka ea bonoa mefuteng e sa sebetseng ea mutant (Liao et al., 2017). Ho hloka UBR5 ho amahanngoa le keketseho ea apoptosis, necrosis, le thibelo ea kholo ea hlahala ho TNBC ka lebaka la angiogenesis e mpe. Ka lebaka la tahlehelo ea UBR5, ho ata ha hlahala ho ea lithong tse hole hoa fokotseha, 'me UBR5 e baka EMT e sa tloaelehang haholo-holo ka ho fokotsa tlhahiso ea E-cadherin (Zhang le Weinberg, 2018). Haufinyane tjena, UBR5 e bontšitsoe e le ntlha ea bohlokoa haholo ho IFN-γ-induced PDL1 transcription ho TNBC ka lebaka la ho hloka ts'ebetso ea eona ea ubiquitination ea E3. Tlhahlobo ea RNA transcriptome e senotse hore UBR5 e ka ba le litlamorao tsa tsamaiso liphatseng tsa lefutso tse amanang le tsela ea IFN-γ le ho khothaletsa transactivation ea PDL1 ka ho eketsa maemo a ts'ebetso ea protheine kinase RNA (PKR) le li-transducer tsa eona tsa matšoao le li-activator. transcription 1 (STAT1) le interferon regulatory factor 1 (IRF1). Leha ho le jwalo, ho hlapollwa ha UBR5 le PD-L1 ho kopantsweng ke CRISPR/Cas9 ho na le phello ya kalafo e sebetsang hammoho ho feta thibelo efe kapa efe feela, ka ditlamorao tse tebileng tikolohong e nyane ya hlahala (Wu et al., 2022). Kahoo, CRISPR/Cas9 e ka ba sesebediswa sa bohlokwa sa ho thibela tshebetso ya UBR5, ka hona ho thibela metastasis ya TNBC le kgolo ya hlahala.
ROR1 ke protheine ea mofuta oa I transmembrane e hlahisoang nakong ea mofetše le nts'etsopele ea lesea le sa tsoa emoloa 'me e fumanoe e le protheine ea oncofetal (Nicholas Borcherding, 2014). Boitšoaro bo hlaselang ba mefuta e fapaneng ea mofetše oa batho bo amahanngoa le ho eketseha ha ROR1. Liphetho tse tšepisang li fumanoe lithutong tsa in vivo le in vitro tse kenyeletsang metsoako ea phekolo e shebaneng le ROR1 (Chien et al., 2016). Maemo a phahameng a ROR1 mRNA libukeng tsa lisele tsa matsoele le 'ona a amahanngoa le lihlahala tse matla tsa matsoele tse kang basal (BL) le ho falla ha tsona likarolong tse ling tsa' mele. Ho feta moo, ho pepeseha ho feteletseng ha ROR1 ho fumanoe e le letšoao la ho bolela esale pele bakeng sa nts'etsopele ea TNBC (Chien et al., 2016). Leha ho le joalo, ho khutsisa ROR1 ho sebelisa CRISPR/Cas9 ho thibela kholo ea TNBC le ho ata ha lisele e tla ba leano le sebetsang.
Ts'ebetso ea sialylation e kenyelletsa ho eketsoa ha sialic acid ho li-glycoconjugates, tse susumetsoang ke sialyltransferases (STs). ST8SIA1 ke ea lelapa la ST mme e bapala karolo ea bohlokoa ho pathogenesis ea mafu a fapaneng a kang leukemia ea lymphocytic le kankere ea mala a maholo (Chang et al., 2018). Tlhahlobo ea tatellano ea RNA e bontša hore ST8SIA1 e hlahisoa haholo liseleng tsa matsoele tsa bakuli ba TNBC 'me e amana hantle le liphetoho tse teng gene ea tumor suppressor p53, e ka kenyang letsoho ho pathogenesis ea TNBC (Battula et al., 2017). Ho phaella moo, ST8SIA1 e ameha ho metastasis le ho khutla ha TNBC, e leng se bontšang hore e bapala karolo ea bohlokoa ho hlaheng le nts'etsopele ea TNBC. Mohlaleng oa in vitro oa TNBC, ho ile ha bontšoa hore ST8SIA1 e theotsoe ke CRIPSR/Cas9 e thibela kholo le metastasis (Battula et al., 2017). Sena se fana ka maikutlo a hore CRISPR/Cas9 e ka ba sesebelisoa sa bohlokoa bakeng sa ho thibela ts'ebetso ea oncogene ea ST8SIA1 kalafong ea TNBC.
NAT1 ke enzyme ea metabolism e susumetsang ho thehoa ha metsoako ea xenobiotic ea karolo ea II 'me e hlahisoa hoo e batlang e le liseleng tsohle tsa motho. NAT1 e ka sebelisa cofactor folate ho shebana le acetyl-CoA (acetyl-CoA) esita le ha ho se na aromatic amine substrate (Stepp et al., 2015; Laurieri et al., 2014). Lithuto li bontšitse hore NAT1 e laola mosebetsi oa matrix metalloproteinase 9 (MMP9) mehlala ea lisele tsa kankere ea matsoele 'me e sireletsa khahlanong le mefuta ea oksijene e arabelang (ROS) nakong ea tlala ea glucose (Wang et al., 2018). Ho tlosoa ha NAT1 ho bontšitsoe ho thibela pyruvate dehydrogenase complex, e leng se lebisang ho se sebetseng hantle ha mitochondrial (Wang L. et al., 2019). Ho phaella moo, litlaleho tse ling tse fapaneng li bontšitse hore thibelo ea NAT1 e sebelisang limolek'hule tse nyane le ho khutsisa siRNA e ka fokotsa ho kenella le ho ata ha lisele tsa kankere ea matsoele (Stepp et al., 2018). Haufinyane tjena, CRISPR/Cas9 e sebelisitsoe ho thibela NAT1 moleng oa sele ea kankere ea matsoele MDA-MB-231, e amang metabolism ea sele ho latela boemo ba eona ba tlhahiso. Phuputso ena e boetse e bontšitse hore NAT-1 e bohlokoa bakeng sa tsoelo-pele le metastasis ea TNBC (Carlisle et al., 2020).
Phetolelo e hokahaneng ea li-oncogene e laoloa ke li-super enhancers, lintlha tsa phetolelo le li-cofactors (Hnisz et al., 2015). Ho phaella moo, sehlopha sa li-kinase tse itšetlehileng ka cyclin (li-CDK), tse kang CDK7, CDK8, CDK9, CDK12, le CDK13, lia hlokahala bakeng sa taolo ea phetolelo. Har'a tsona, CDK7 e ameha ho phosphorylation ea RNA polymerase II, e leng ea bohlokoa haholo bakeng sa ho qalisoa le ho atolosoa ha phetolelo ea oncogene ho pathogenesis ea TNBC. Phuputsong ea bohlokoa, ho tlosoa ha CDK7 ke CRISPR/Cas9 ho ile ha thibela TNBC, ho bontšang hore pathogenesis ea TNBC e itšetlehile ka CDK7 (Wang Y. et al., 2015).
Lithuto li bontšitse hore liphetoho tsa MYC le RBP (protheine e tlamang ea RNA) li ka lebisa ho apoptosis, ha liphetoho tsa liphatsa tsa lefutso tse le 'ngoe ho MYC kapa RBP li sa ame kholo ea lisele tsa mofetše (Einstein et al., 2021). Li-RBP tse fetang 1000 ho genome ea motho li hlahlobiloe ho sebelisoa laeborari e thehiloeng ho CRISPR/Cas9. Har'a tsona, li-RBP tse 57 li fumanoe li le bohlokoa bakeng sa kholo ea lisele tsa mofetše tse nang le maemo a phahameng a MYC (Wheeler et al., 2020). Ho phaella moo, YTHDF2 e bohlokoa bakeng sa ho boloka kholo ea lisele tsa TNBC 'me e fokotsa palo ea lingoloa tse nang le methylated nakong ea phetolelo ea boemo bo holimo liseleng tsa mofetše tse nang le tlhahiso e phahameng ea MYC. Ho feta moo, YTHDF2 ha e bohlokoa bakeng sa lisele tsa mofetše, tse sa itšetleheng haholo ka maemo a phahameng a MYC ho lelefatsa ho phela ha bakuli ba TNBC (Einstein et al., 2021), e leng se bontšang hore e ka ba sepheo se ka bang teng sa kalafo bakeng sa meriana e ka hlōlang TNBC.
Liprotheine tsa menoana ea zinki (ZNFs) li etsa hoo e ka bang 1% ea genome eohle ea motho. Lithuto li bontšitse hore ZNF e ka laola ho ata ha lisele mefuteng e fapaneng ea kankere, joalo ka kankere ea sebete, kankere ea matsoele le kankere ea mala a maholo (Zhang W. et al., 2021; Li et al., 2017). Lilaeborari tse hlahisoang ke CRISPR knockout li sebelisitsoe ho hlahloba liphatsa tse fapaneng tsa ho thibela hlahala (TSGs) liseleng tsa kankere ea matsoele (Shalem et al., 2014). Ho tloha ka nako eo, phuputso ea transcriptomic ea lisele tsa kankere ea matsoele tse hlakotsoeng tsa CRISPR/Cas9 ZNF319 e bontšitse hore ZNF319 ke liphatsa tsa lefutso tsa ho thibela hlahala tse fokotsang ho ata ha kankere ea matsoele 'me ka hona li ameha mekhoeng e fapaneng ea lipontšo le mesebetsing e meng ea baeloji (Wang L. et al., 2022).
Ferroptosis ke mofuta oa lefu la sele le hlophisitsoeng le itšetlehileng ka boteng ba tšepe. Hoa tsebahala hore nts'etsopele ea ferroptosis e susumetsoa ke boteng ba lipid peroxides. Ho phaella moo, PKCβII e tlalehiloe e bontša peroxidation ea lipid pele, 'me keketseho ea peroxidation ea lipid e amahanngoa le ferroptosis. Ho hlahlojoa ha laeborari ea li-kinase inhibitors tse sebetsanang le CRISPR/Cas9 ho senotse hore PKCβII e ameha ts'ebetsong ea peroxidation ea lipid, e leng ea bohlokoa bakeng sa ferroptosis liseleng tsa MDA-MB-231 (Zhang et al., 2022). Ka hona, sena se bontša hore ho tlosoa ha PKCβII ke CRISPR/Cas9 e ka ba sepheo se ka bang teng sa ho thibela hlahala bakeng sa kalafo ea mafu a amanang le ferroptosis (Zhang et al., 2022).
Ho lumeloa hore ho hanyetsa lithethefatsi ho ikarabella bakeng sa hoo e ka bang 90% ea mafu ho bakuli ba kankere 'me ke e' ngoe ea liphephetso tse kholo kalafong ea kankere (Bukowski et al., 2020). Liphetho li bontša hore palo e lekaneng ea liphatsa tsa lefutso tse amanang le ho tsoa ha lithethefatsi, tokiso ea DNA, apoptosis le litsela tse fapaneng tsa lipontšo tsa sele li amahanngoa le ho hanyetsa lithethefatsi (Haider et al., 2020). Har'a tsona, liphatsa tsa lefutso tse 'maloa li shebiloe ke lisebelisoa tsa CRISPR/Cas9' me li bontšitse liphetho tse tšepisang ho fokotsa ho hanyetsa lithethefatsi le ho eketsa katleho ea mekhoa ea phekolo ea kankere (Vaghari-Tabari et al., 2022). Ho phaella moo, lilaebrari tsa tlhahlobo ea liphatsa tsa lefutso tsa CRISPR/Cas9 tse nang le tšusumetso e phahameng li 'nile tsa ameha ho fetoleng mosebetsi oa lipheo tse ka bang teng tsa liphatsa tsa lefutso tsa ho hanyetsa lithethefatsi (Shalem et al., 2015). Ho hlwaya dijini tsa ho hanyetsa paclitaxel, ho sebediswa tatellano ya RNA e tsamaellanang le tlhahlobo ya laeborari ya sgRNA ya genome ka bophara ho hlwaya dijini tse robedi tse ka bang teng, ho kenyeletswa le histone deacetylase 9 (HDAC9), tse amanang le ho hanyetsa dithethefatsi ho bakudi ba nang le TNBC e iphetang (B et al., 2020). ). Phuputsong e nngwe, ho bonwa keketseho ya tlhahiso ya diserine/threonine le tyrosine protein kinase (DSTYK) nakong ya ho phela ha bakudi ba TNBC ba phekolwang ka meriana e thibelang kankere. Ho feta moo, ho kgutsufala ha DSTYK ho bakwang ke CRISPR/Cas9 ho ntlafaditse haholo apoptosis ya disele tsa kankere tse hanyetsanang le dithethefatsi ka hara vitro (disele tsa SUM102PT le disele tsa MDA-MB-468) le mohlala wa TNBC ka hara vivo (Ogbu et al., 2021).
Leha TNBC e na le ts'ebetso e sa tloaelehang ea tsela ea MAPK, phello ea kliniki ea mekhoa ea phekolo e shebaneng le MEK e mpe. Sekero sa laeborari ea genomic ea CRISPR/Cas9 se senotse hore thibelo ea PSMG2 (proteome assembly chaperone 2) e etsa hore lisele tsa TNBC BT549 le MB468 li utloahale ho MEK inhibitor AZD6244. Ho oa ha PSMG2 ho CRISPR/Cas9 ho fetotse ts'ebetso e tloaelehileng ea proteasome, e leng se ileng sa lebisa ho qhalaneng ha PDPK1 ka autophagy, ka hona ho ntlafatsa lisele tsa hlahala ho mehlala ea litoeba tsa TNBC e hlahisitsoeng ke AZD6244 (MEK inhibitor) le MG132 (proteasome inhibitor). Kahoo, li-inhibitor tsa proteasome le MAP kinase (MEK) li ka fanoa ka kopanelo ho fokotsa ho ata ha lisele tsa hlahala (Wang X. et al., 2022).
CRISPR/Cas9 e boetse e sebelisitsoe ho hlahloba tahlehelo ea ts'ebetso ea liphatsa tsa lefutso e lebisang ho khanyetso ea TNBC (Shu et al., 2020). Ge et al. ba hlalositse mokhoa oa khanyetso ea lithethefatsi ea lisele tsa mofetše tse phekoloang ka JQ1, BET bromodomain inhibitor (BBDI), ho TNBC. Ba sebelisa CRISPR/Cas9, ba fumane hore ho hlakola liphatsa tsa lefutso tsa rb1 ho bakile hore TNBC e hane ho moriana o loantšang mofetše JQ1. Ka hona, ts'ebetso ea rb1 e bohlokoa karabelong ea meriana ea JQ1 ho TNBC. Ba boetse ba tlaleha hore paclitaxel ke inhibitor ea CDK4/6 kinase/microtubule, 'me motsoako oa eona le li-BBDI tse kang JQ1 o ka fana ka likarabo tse tšepisang tsa kalafo ho TNBC e hanyetsanang le meriana (Ge et al., 2020).
Sebaka sa ho ngolwa ha RNA e telele e sa ngoleng khoutu (lncRNA) se tlalehiloe se ikarabella bakeng sa ho hanyetsa kalafo ea neoadjuvant ho TNBC. Phuputso ena e bontša hore lingoloa tse hlano tse fapaneng tsa MALAT1 lncRNA li hlahisoa haholo ho TNBC. Ho phaella moo, ho tlosoa ha MALAT1 ka thuso ea CRISPR/Cas9 ho ekelitse kutloisiso ea lisele tsa TNBC BT-549 ho paclitaxel le doxorubicin, e leng se bontšang karolo e ka bang teng bakeng sa ho hanyetsa MALAT1 ho TNBC (Shaath et al., 2021).
Liphetoho tsa lefutso ho BRCA1 (BRCA1m) lia fapana 'me ka hona ho thata ho li lemoha. Ho shebana le PARP1 (poly(ADP-ribose) polymerase), molekane ea bolaeang oa maiketsetso oa BRCA1, ho ka eketsa kutloisiso ea chemo ea meriana ea TNBC. Ho sebelisa CRISPR/Cas9, ho tlosoa ha PARP1 ho ekelitse kutloisiso ea meriana e loantšang mofetše joalo ka doxorubicin, gemcitabine le docetaxel ho lisele tsa TNBC tse fetotsoeng tsa mBRCA1, ho fana ka maikutlo a hore PARP1 le eona e ameha ho hanyetseng lithethefatsi ho TNBC (Vaghari-Tabari et al., 2022). Liphetoho tsa lefutso tse hatellang hlahala BRCA1 kapa BRCA2 li tsejoa li eketsa monyetla oa ho ba le mofetše oa matsoele ho batho. Kalafo ea bakuli bana e hloka tšebeliso ea li-inhibitor tsa PARP. Ho phaella moo, ho bontshitswe hore ho kgaolwa ha nucleotide salvage factor DNPH1 ho sebedisa CRISPR/Cas9 ho ka tlosa nucleotide e nang le chefo ya 5-hydroxymethyldeoxyuridine (hmdU) monophosphate, ka hona ho ntlafatsa karabelo ya disele tse haellang BRCA ho ba le kutloelo-bohloko ho di-inhibitor tsa PARP (Fugger). et al., 2021). Kahoo, di-inhibitor tsa CRISPR/Cas9 le PARP1 e ka ba leano la bohlokwa la kalafo bakeng sa TNBC.
Jini ea penetrant glycoprotein (P-gp) ke jini ea ho hanyetsa meriana e mengata eo ka kakaretso e eketsehang ho hoo e ka bang 41% ea TNBC eohle (Sun et al., 2020). Ho tsoa ha meriana e bakoang ke P-gp ho fumanoe e le molaoli oa bohlokoa oa ho hanyetsa meriana kankereng ea matsoele. Lithibela-P-gp li bontšitse kutloisiso e eketsehileng ea meriana e loantšang mofetše kankereng ea matsoele (Famta et al., 2021). Kahoo, ho tlosoa kapa ho hatelloa ha lithibela-P-gp le P-gp tse tsamaisoang ke CRISPR/Cas9 e ka ba mekhoa ea bohlokoa haholo ea ho hlola ho hanyetsa meriana ho TNBC.
Sepalami sa khasete se tlamang ATP G2 (ABCG2) se tsebahala ka ho baka ho hanyetsa lithethefatsi ho TNBC (Palasuberniam et al., 2015), leha ho le jwalo ha ho na ditlaleho mabapi le kamano pakeng tsa ho kgutsa ha CRISPR/Cas9 le ABCG2 le ho se sebetse ha gene e thibelang hlahala. PTEN E ka eketsa ho sebetsa ha ABCG2 (Palasuberniam et al., 2015), (Deepak Singh et al., 2021). Ka hona, ho sebedisa CRISP/Ca9 ho tlosa ABCG2 le ho e kopanya le sethibela-mafu sa ABCG2 e ka ba kalafo e loketseng ho hlola ho hanyetsa lithethefatsi ho TNBC. Tafole ya 2 e bua ka CRISPR/Cas9 e shebaneng le dijini tsohle tse hanyetsanang.
Tafole ea 2. CRISPR/Cas9 e shebane le liphatsa tse fapaneng tsa lefutso tsa ho hanyetsa TNBC ho susumetsa lisele ho ea merianang e thibelang kankere.
Lilaeborari tsa CRISPR/Cas9 ke lisebelisoa tse ka bang teng bakeng sa ho hlahloba liphetoho tsa liphatsa tsa lefutso tse amanang le pathogenesis ea kankere (Chan et al., 2022). Mokhoa ona oa ho hlahloba o kenyelletsa mehato e mene e kang (a) kaho ea laeborari, (b) phetisetso ea lentiviral, (c) tlhahlobo ea phenotypic, le (d) tlhahlobo ea liphatsa tsa lefutso tse reretsoeng. Leha TNBC e na le ts'ebetso e sa tloaelehang ea tsela ea MAPK, tebello ea kliniki ea phekolo e reretsoeng MEK bakeng sa bakuli ba TNBC ba jereng liphetoho tsa liphatsa tsa lefutso tse hatellang hlahala tse kang PTEN, RB1, le TP53 e fokola haholo.
Ho phaella moo, tlhahlobo ya ho kgaotsa dijini e sebedisang CRISPR/Cas9 e lekile molek'hule e matla le e kgethilweng ka ho fetisisa, dehydrocatrol, e nang le di-extract tsa dipalesa le mahlaku a Guatemala, ho utlwisisa phello ya dehydrocatrol ho MDA-MB-231. Mekgwa ya ho tjhesa disele tse kgethilweng. Dibopeho tsa Mesenchymal stemness tsa mefuta e fapaneng ya TNBC. Sekerine sena se thehilweng ho CRISPR/Cas9 se boetse se senotse hore HSD17B11, jini e kenyang 17β-hydroxysteroid dehydrogenase type 11, e hlahiswa haholo diseleng tsa MDA-MB-231 mme e fella ka dehydrofalcarinol e ikgethang bakeng sa disele tsa MDA-MB-231 (Grant et. al., 2020). Kahoo, sena se fana ka maikutlo a hore tlhahlobo ya genomic ya CRISPR/Cas9 e na le bokgoni bo boholo ba ho hlwaya mekgwa e itshetlehileng hodima thepa ya ho thibela kankere ya metswako ya tlhaho e ka bang teng.
Ho phaella moo, ho ba kotsing ha TNBC ho kankere ho ithutilwe ho sebediswa skrine ya CRISPR/Cas9 e sa nke lehlakore, e bontshitseng kamano pakeng tsa ditsela tsa oncogenic le tse hatellang hlahala. Dikarolo tsa bohlokwa tsa ditsela tsa mTOR le Hippo di tlalehilwe di bapala karolo ya bohlokwa taolong ya hlahala ho TNBC. Ho feta moo, dithuto di bontshitse hore thibelo ya meriana ya mTORC1/2 le YAP oncoprotein e thibela ka katleho ho baka tshwaetso ya TNBC ho sebediswa mohlala wa ho kopanya meriana le matrix ka hara vitro le di-xenograft tse tswang ho mokuli ka hara vivo. Ho feta moo, thibelo ya mTORC1/2 e tsamaiswang ke Torin-1 e eketsa macropinocytosis, athe thibelo ya YAP e bakwang ke vertebrorfin e lebisa lefung la sele ya TNBC. Ha di kopantswe, diphetho tsena di totobatsa matla le matla a tlhahlobo ya CRISPR ya genome ka hara vivo ho hlwaya kalafo e ntjha le e sebetsang bakeng sa TNBC (Dai et al., 2021).
Ho se sebetse hantle ha sesole sa mmele ke ntlha ya bohlokwa ho qaleng ha hlahala. Disele tsa mofetshe di qoba ho hlweka ha sesole sa mmele ka ho qoba mekgwa ya tshireletso, ho kenyeletswa le ho sitisa tshebetso ya disele tsa mmele tsa mmele tikolohong e nyane ya hlahala le ho beha sesole sa mmele kotsing. Ka hona, ho ntshetsa pele tsamaiso e ntlafetseng ya mmele ya ho itšireletsa mafung e ka ba mokgwa wa bohlokwa wa ho lwantsha dihlahala. Tshebediso ya phetoho ya lefutso e thehilweng ho CRISPR/Cas9 e sebetsana le mathata a mmalwa a amanang le ho se sebetse hantle ha sesole sa mmele ho ya ka maikutlo a fapaneng. CRISPR/Cas9 e sebediseditswe ho ntlafatsa sesole sa mmele sa ho lwantsha hlahala kgahlanong le kankere ya matswele ka ditsela tse latelang (Setshwantsho sa 3).
Setšoantšo sa 3. Kalafo ea 'mele ea CRISPR/Cas9 e shebane le lisele tsa TNBC. Ho lahleheloa ke CDK5 le ho oa ha PDL1 le CD155 ho ntlafatsa sesole sa 'mele. Ka mokhoa o ts'oanang, ho lahleheloa ke A2AR le ho oa ha li-TAA tse kang HER2, mucin 1 le TEM8 ho ekelitse katleho ea lisele tsa CAR-T ho bolaeeng lisele tsa mofetše. Tlhahlobo ea lisele tsa T tse tsamaisoang ke CRISPR/Cas9 e bontšitse hore ho sitisoa ha p38 kinase ho ntlafatsa ts'ebetso ea antitumor ea sele ea T. Lisele tsa T tse fetotsoeng ke CRISPR/Cas9 li eketsa tlhahiso ea TCR (receptor ea sele ea T), e leng se fellang ka hore lisele tsa T li be le ts'ebetso ea antitumor.
Sepheo sa kalafo ea 'mele ke ho hlasimolla sesole sa 'mele ho hlasela lisele tsa mofetše. Ho ata ha liprotheine tsa tlhahlobo ea 'mele hangata ho thibela liketso tsa 'mele tse itšireletsang empa ho ka thusa mofetše ho li qoba (Topalian et al., 2015). Liprotheine tsena li thibela liketso tsa 'mele ka ho tlama li-receptor holim'a lisele tsa 'mele tse itšireletsang. Liprotheine tse ngata tsa tlhahlobo (tse kang CD155 le PD-L1) li shebana le receptor ea PD-1 liseleng tsa 'mele tse itšireletsang 'me li hlahisoa mofetšeng oa matsoele, haholo-holo TNBC (Li Y.-C. et al., 2020). Ho theola PD-L1 kapa receptor ea eona ho sebelisa CRISPR/Cas9 ho ka hlasimolla sesole sa 'mele ho hlasela lihlahala tsa TNBC (Yahata et al., 2019). Ho phaella moo, ho fokotseha ha polelo ea PD-L1 ka ho tlosoa ha CDK5 ke CRISPR/Cas9 ho bontšitsoe ho thibela kholo ea hlahala ka har'a vitro le ka har'a 'mele (Deng et al., 2020). Thibelo ea kholo ea kankere ea matsoele ka har'a mehlala ea in vitro le in vivo e bontša hore phokotso ea CD155 e tsamaisoang ke shRNA e ka ba le phello ea phekolo mofetšeng oa matsoele (Gao et al., 2018).
Lisele tsa CAR T li hlahisa li-CAR tse lemohang li-antigen tse amanang le hlahala (TAAs) 'me li ka sebelisa ho theola liprotheine tsa checkpoint ho ntlafatsa ts'ebetso ea tsona (Li C. et al., 2020). Ho na le lipheo tse fapaneng tse ka bang teng bakeng sa kalafo ea lisele tsa CAR T mofetšeng oa matsoele, ho kenyeletsoa li-TAA tse 'maloa tse kang HER2, mucin1, le TEM8 (Bajgain et al., 2018). Ho na le bopaki ba hore lisele tsa CAR T tse shebaneng le mesothelin (e hlahisoang haholo liseleng tsa TNBC BT-459) li sebetsa hantle khahlanong le mofetše ha PD-1 e tlosoa ho sebelisoa CRISPR/Cas9 (Hu et al., 2019). Leha ho le joalo, ho arola lisele tsa T ho bakuli ebe u li hlophisa ex vivo ke ts'ebetso e boima le e jang nako. Le hoja lisele tsa T tse akaretsang li ka fokotsa litlhoko tsa ho itšehla thajana, lisele tsa T tsa mofani tse hlahisang human leukocyte antigen (HLA) class I le T cell receptor (TCR) li lokela ho tlosoa ho thibela ho se tloaelehe ha graft-versus-host (Ren et al., 2017). , 2017a). Ho sebelisoa CRISPR/Cas9, HDR e ka felisa TCR le HLA ka nako e le 'ngoe le ho tlosa CAR e kenyang liphatsa tsa lefutso. Lithuto li bontšitse hore CRISPR/Cas9 e ka sebelisoa ho kenya li-CAR tse khahlanong le CD19 sebakeng sa TCR, ka hona e hlahisa li-CAR ka katleho ntle le ho felisa lisele tsa T (Dimitri et al., 2022). Mahlale a mangata a ntlafalitsoe ho hlahisa lisele tsa CAR T tse nang le allogeneic ka ho felisa TCR, beta-2-microglobulin (B2M), HLA-I subunit le liprotheine tse ling tse kang PD-1 le CTLA-4, tse ka bang le thepa e kholo ea ho loantša kankere khahlanong le TNBC (Eyquem et al., 2017; Liu et al., 2017; Ren et al., 2017b; Dimitri et al., 2022).
Ho sebelisa CRISPR/Cas9 ho ka ntlafatsa katleho ea lisele tsa CAR-T. Adenosine e tsejoa e na le thepa ea ho thibela sesole sa 'mele' me e ka fokotsa tšireletso ea 'mele khahlanong le mofetše ka ho thibela ts'ebetso ea lisele tsa T le ho kenya tšebetsong li-receptor tsa adenosine A2A (A2AR) (Vigano et al., 2019). Ho khutsisa A2AR ho sebelisa CRISPR/Cas9 ho ntlafalitse haholo katleho ea lisele tsa CAR-T ka har'a 'mele (Giuffrida et al., 2021). Lisele tsa T li tsejoa ka ho bontša litšobotsi tse fapaneng tsa phenotypic tse kang katoloso ea sele, phapang, khatello ea oxidative le khatello ea genomic. Tlhahlobo ea lisele tsa T tse thehiloeng ho CRISPR/Cas9 e senotse tšitiso ea li-kinase tse 25 tse fapaneng tsa T cell receptor. Har'a tsona, ho tlosoa ha p38 kinase ho bontšitsoe ho ntlafatsa ts'ebetso ea antitumor ea lisele tsa T, ho bontšang hore p38 kinase ke molaoli oa bohlokoa oa taolo ea lisele tsa CAR-T (Gurasamy et al., 2020).
Lisele tsa T li ka fetoloa liphatsa tsa lefutso ho sebelisoa CRISPR/Cas9 ho hlahisa li-TCR tse hlahisoang haholo. Ho fetisetsa lisele tsa T tse fetotsoeng liphatsa tsa lefutso ho bakuli ho bontšitse ts'ebetso e matla ea ho loantša mofetše ho feta lisele tsa T tse sa tloaelehang. Ka hona, li-TCR tse sa tloaelehang li ka qothisana lehlokoa le li-TCR tse fetotsoeng liphatsa tsa lefutso ho bakuli, e leng se ka amang bokhoni ba kalafo ea mofetše. Ho hlōla bothata bona, ho sebelisa CRISPR/Cas9 ho tlosa TCR-β e sa tloaelehang liseleng tse amohelang le ho fetisetsa lisele tsa T tsa T tsa T ho bakuli ba mofetše ho ka bontša likarabo tse betere tsa 'mele khahlanong le mofetše ntle le tlhoko ea tlholisano ea thobalano ea TCR e sa tloaelehang (Fan et al. 2018). Kahoo, ntle le lisele tsa T tse fetotsoeng tsa CRISPR, li-TCR li na le kutloelo-bohloko e fetang makhetlo a sekete ho li-antigen tsa hlahala ho feta lisele tse tloaelehileng tsa T tse fetotsoeng ke TCR. Ho feta moo, ho li-leukemia tse fapaneng, lisele tsa T tse fetotsoeng tse hlahisoang ke γδ TCR+ CRISPR li bontša tlhahiso e phahameng ea lisele tsa T tsa CD4+ le CD8+ ho feta phetiso e tloaelehileng ea TCR (Legut et al., 2018). Ka hona, lisele tsa T tse fetotsoeng tse hlahisoang ke CRISPR/Cas9 e ka ba mokhoa o atlehang oa phekolo ea immunotherapy ho hlola TNBC.
Li-Integrin ke limolek'hule tsa ho khomarela lisele tse teng ka har'a li-transmembrane tsa sele 'me li khothalletsa ho tlama ha lisele ho matrix ea kantle ho sele (ECM) (Hamidi le Ivaska, 2018). Ho se sebetse hantle ha li-integrin ho amahanngoa le nts'etsopele le ho falla ha kankere ka ho fetola matrix ea kantle ho sele, e leng se lebisang ho pholoheng ha lisele tsa kankere potolohong ea mali (Hamidi le Ivaska, 2018). Ho kokota ha integrin ho tsamaisoang ke CRISPR/Cas9 ho liehisa tsoelo-pele ea hlahala, metastasis le colonization ho TNBC. Ho kokota ha integrin a5 (ITGA5) ho tlalehiloe ho fokotsa ho falla ha lisele le tsoelo-pele ho kankere tse ling tse kang kankere ea matšoafo (Ju et al., 2017), ho fana ka maikutlo a hore integrin a5 e ka boela ea e-ba ntlha ea bohlokoa ho pathogenesis ea TNBC.
Ho hlahisa litoeba tse thibaneng ka ho sebelisa mekhoa ea setso ea lisele tsa embryonic stem (ES) ke ts'ebetso e boima, e jang nako, le e sa sebetseng hantle. Ho nka likhoeli le lilemo ho shebana le lisele tsa ES ka ho kopanya litoeba tse tšoanang, ho hlahisa litoeba tsa chimeric, ebe li li kopanya le litoeba tse nang le li-heterozygous ho hlahisa bana ba nang le li-homozygous. Ho hlokahala ho kopanya litoeba ka liphetoho tse ngata tsa lefutso. Leha ho le joalo, ho hlahile mathata a mangata ha ho sebelisoa litoeba tse hlahisoang ke ho tlosoa ha lisele tsa ES ho sebelisoa CRISPR/Cas9 kapa ka ho kenya likarolo tsa CRISPR/Cas9 ka har'a mahe a nontšitsoeng a sele e le 'ngoe. Mohlala, ho hlahisoa ha liphetoho hangata ho etsahala sebakeng sa biallelic 'me ha ho na liphatsa tsa lefutso. Ho sa tsoa tlalehoa hore lisele tsa ES tse sebelisang theknoloji ea CRISPR/Cas9 li ka kenya liphetoho tsa biallelic ka nako e le 'ngoe liphatseng tsa lefutso tse fihlang ho tse 5 (Nishizono et al., 2021). Bakeng sa morero ona, Cas9 mRNA le li-gRNA tse hlano tse ikhethileng tsa liphatsa tsa lefutso li ile tsa fetisetsoa ka nako e le 'ngoe liseleng tsa ES. Sena se totobatsa tšepiso le katleho ea mokhoa ona, leha liphetoho tsena li ka be li fetiselitsoe ha mela ea mothehi e ne e hlahisoa ho hlahisa litoeba tse hlano tse kobehileng. Phuputso ena e boetse e tlaleha tšibollo e makatsang: Lisele tsa ES ha li sa hlokahala ho theha litoeba tse fetotsoeng liphatsa tsa lefutso. Ho e-na le hoo, ho tlosa lihlahisoa tse itseng tsa liphatsa tsa lefutso, Cas9 mRNA le gRNA li ile tsa kenngoa ka har'a li-embryo tsa sethala sa sele e le 'ngoe. Ka lebaka leo, mela ea mothehi ea kobehileng e entsoe e ka sebelisoang ka khopolo-taba ho ithuta liphello tsa ho tlosoa ha liphatsa tsa lefutso ho litoeba (Qin et al., 2016). Kahoo, CRISPR/Cas9 e lumella ho thehoa ha litoeba tse fetotsoeng liphatsa tsa lefutso ho phekola TNBC ka litšenyehelo tse tlase ho feta boenjiniere ba liphatsa tsa lefutso ba setso. Ho sebelisoa sistimi ea CRISPR/Cas, mohlala o mocha oa mothehi oa kobehileng o ntlafalitsoe o ka atlehang ho hlahisa liphetoho tsa lintlha ho liphatsa tsa lefutso tse le 'ngoe kapa tse ngata tsa tlhaho ho TNBC. Ho latela mohopolo ona, mehlala ea liphoofolo ea TNBC e ka boela ea ntlafatsoa ka ho sebelisa ho tlosoa ha BRCA1 le p53 ka thuso ea CRISPR/Cas9, e leng se fellang ka tahlehelo ea tokiso ea HR, ho se tsitse ha genomic, le li-phenotype tse fetotsoeng (Annunziato et al., 2020).
Hona jwale, mekgwa e fapaneng e kang mammography, magnetic resonance imaging (MRI) le ultrasound e sebediswa ho hlahloba TNBC. Leha ho le jwalo, mekgwa ena e na le mefokolo e itseng. Mammography e sebediswa ho hlahloba dinama tsa matswele tsa sebakeng seo ho ena le metastases, moo disele tsa mofetshe di falletseng dibakeng tse ding. Ultrasonography ha se mokgwa o tshepahalang wa ho hlahloba TNBC (Chen le Lee-Felker, 2023). MRI e na le kutlwisiso e phahameng ho feta ultrasound le mammography, empa ho nepahala ha yona ha tlhahlobo ho lekanyeditswe (Sha le Chen, 2022). Biopsy ya dinama ke tsela e hlaselang ya ho lemoha disele tsa mofetshe. Leha ho le jwalo, maemong a mang, biopsy e ka fosa dinama tsa mofetshe haeba nale e kheloha sebakeng seo e se ratang. Ho feta moo, e theko e boima haholo mme e ka ba kotsi ho mokuli. TNBC ke mofuta o fapaneng wa mofetshe, kahoo biopsy e kanna ya se fane ka tlhahisoleseding e lekaneng mabapi le mofuta wa mofetshe. Ka hona, mokgwa o motjha o tshepahalang wa tlhahlobo ya ho lemoha TNBC o hlokahala ka potlako. CRISPR/Cas9 e ka sebetsa e le mokhoa o mong o utlwahalang haholo le o sa keneleng haholo bakeng sa tlhahlobo ya mofetshe wa matswele. Ho fihlela sena, maano a thehiloeng ho CRISPR/Cas9 a ka sebediswa ho ntlafatsa mekgwa ya PCR bakeng sa ho fumana TNBC. Taba ya pele, diprotheine tsa Cas9 le cpf1 tsa sistimi ya CRISPR di sebediswa ho tlosa DNA e seng e ikgethang, mme diprotheine tsena tse pedi (Cas9 le cpf1) di ka lemoha tatellano ya PAM pele di tlamahana le DNA e rerilweng (Deepak Singh et al., 2021). . Kahoo, PCR e ka hlwaya diphetoho tse amanang le ntshetsopele ya kankere. Diphuputso tse mmalwa di amohetse leano lena le thehilweng ho CRISPR ho lemoha diphetoho tse fapaneng dikankereng tse fapaneng (Safari et al., 2019). Kahoo, mekgwa ya PCR e thehilweng ho CRISPR e ka fokotsa ho itshetleha ha tlhahlobo ya TNBC mekgweng e hlaselang jwalo ka immunohistochemistry e thehilweng ho biopsy.
Liphetoho tsa genotypic taolong ea li-receptor tsa lihormone le tsona li bontšitsoe ho TNBC (Chen le Russo, 2009). Maano a PCR a thehiloeng ho CRISPR/Cas9 a sebelisang li-microarray bakeng sa tlhahlobo ea libaka tsa tlhokomelo a ka beha leihlo liphetoho tsena ka katleho (Hajian et al., 2019). E ka boela ea fa bafani ba tlhokomelo ea bophelo tlhahisoleseling e nepahetseng mabapi le liphetoho tse itseng ho bakuli ba TNBC, e leng se ka thusang ho ntlafatsa tsela ea bona ea kalafo. Leha ho le joalo, mokhoa ona o kopaneng o na le meeli. Ka hona, ho hlokahala boiteko bo boholo ba lipatlisiso pele mokhoa ona oa CRISPR/Cas-PCR o ka sebelisoa tlhokomelong ea bophelo bakeng sa tlhahlobo ea TNBC (Yang et al., 2019).
Nako ea poso: Mphalane-14-2024