Kev Hloov Pauv Hloov Gene: Kev Nce Qib Hauv CRISPR/Cas Technology Hauv Biomedical Applications
CRISPR/Cas thev naus laus zis, thawj zaug pom tias yog lub cev tiv thaiv kab mob hauv prokaryotes kom paub thiab txiav cov nucleic acids uas nkag mus rau hauv, tau hloov zuj zus mus ua ib lub cuab yeej muaj zog rau kev kho cov noob caj noob ces hauv cov hlwb tsiaj txhu. Cov protein SpCas9, tau los ntawm Cov kab mob Streptococcus pyogenes, yog thawj Cas protein rov siv dua rau kev kho kom raug genome, vam khom cov lus qhia RNA (sgRNA) los tsom mus rau cov kab ke nucleotide tshwj xeeb hauv genome.
Tus sgRNA muaj ib qho scaffold sequence uas tsim nyog rau Cas9 binding thiab ib qho customizable 20-nucleotide spacer uas txhais tau lub hom phiaj genomic. Rau kev ua haujlwm zoo ob-strand breaks (DSB), qhov spacer sequence yuav tsum ua tau raws li ob qho kev cai: nws yuav tsum tsis txhob ua ke nrog lwm cov genomic DNA thiab yuav tsum nyob ib sab ntawm ib qho protospacer adjacent motif (PAM) sequence, uas yog qhov tseem ceeb rau Cas9 binding.
Qhov kev ua haujlwm txiav ob zaug ntawm Cas9 vam khom nws ob qhov chaw nquag, RuvC thiab HNH, uas tso cai rau kev tsim DSB kom meej. Cov kws tshawb fawb tau tsim ntau yam kev siv siv CRISPR/Cas system, suav nrog:
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Kev Tshuaj Xyuas Tag Nrho Genome Knockout: Los ntawm kev tsim ib lub tsev qiv ntawv sgRNA uas muaj txhij txhua yam thiab siv lub siab xaiv, cov kws tshawb fawb tuaj yeem txheeb xyuas cov noob caj noob ces tseem ceeb rau kev ciaj sia ntawm lub cell nyob rau hauv ntau yam xwm txheej.
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Kev Ua Haujlwm thiab Kev Txwv Gene: Siv dCas9, uas tsis muaj kev ua haujlwm ntawm nuclease, cov kws tshawb fawb tuaj yeem tswj kev qhia tawm ntawm cov noob caj noob ces los ntawm kev sib xyaw dCas9 nrog cov tshuaj transcriptional activators lossis repressors, ua rau muaj kev tshawb nrhiav ntxaws ntxaws txog kev ua haujlwm ntawm cov noob caj noob ces.
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Kev Kho Mob Epigenetic: dCas9 tuaj yeem tsim los hloov kho DNA lossis histones, uas tso cai rau kev hloov pauv epigenetic uas tuaj yeem ua rau nce lossis ntsiag to kev qhia tawm ntawm cov noob caj noob ces.
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Kev Tshuaj Xyuas Chromatin: Los ntawm kev sib xyaw dCas9 nrog cov protein fluorescent, cov kws tshawb fawb tuaj yeem pom cov chaw tshwj xeeb hauv cov hlwb uas muaj sia nyob, txhim kho peb txoj kev nkag siab txog chromatin dynamics.
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Kev Txheeb Xyuas Cov Protein Sib Txuas DNA/RNA: Kev sib txuas CRISPR / Cas nrog cov thev naus laus zis sib xws ua rau muaj kev txheeb xyuas cov protein cuam tshuam nrog cov cheeb tsam genomic tshwj xeeb.
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Kev Hloov Pauv ntawm Cov Tshuaj: Lub kaw lus no tuaj yeem ua rau muaj kev hloov pauv hauv cov kab ke DNA uas tau tsom mus rau, ua rau kev kawm txog kev ua haujlwm ntawm cov noob caj noob ces nyob rau hauv kev xaiv siab.
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Kev Ntxig Los Ntawm Transposon: Los ntawm kev sib txuas cov Cas protein nrog transposases, cov kws tshawb fawb tuaj yeem ua tiav kev koom ua ke ntawm exogenous DNA rau hauv genome.
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Kev Tshem Tawm Cov Fragment Loj: Lub kaw lus CRISPR-Cas3 tso cai rau kev tshem tawm cov kab ke DNA loj, muab cov kev kho mob tshiab rau cov kab mob viral.
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Kev Khaws Cia Nco: Cov gRNAs uas tsom mus rau tus kheej tuaj yeem tsim cov kev hloov pauv mus tas li uas ua haujlwm ua cov cim qhia txog kab mob rau cov xwm txheej tshwj xeeb, ua rau txoj hauv kev rau kev taug qab cov lus teb ntawm tes.
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Kev Tshawb Pom Kab Mob: Cov txheej txheem CRISPR/Cas tau siv tau zoo hauv kev kuaj pom cov kab mob sai, suav nrog HIV thiab SARS-CoV-2, txhim kho kev muaj peev xwm kuaj mob.
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Kev Tsim Kho Tshiab Txog Kev Kho Mob Gene: Qhov kev qhia txog thawj cov tshuaj kho noob caj noob ces uas siv CRISPR/Cas9, CASGEVY™, yog ib qho tseem ceeb hauv kev kho mob sickle cell disease (SCD) thiab transfusion-dependent β-thalassemia (TDT). Cov tshuaj no, uas tau pom zoo hauv ntau thaj chaw, qhia txog lub peev xwm ntawm kev kho noob caj noob ces los hloov pauv cov kev kho mob.
Raws li thaj chaw ntawm kev kho cov noob caj noob ces txuas ntxiv mus, cov ntawv thov ntawm CRISPR / Cas thev naus laus zis tab tom nthuav dav sai. Nrog rau kev nqis peev tseem ceeb ntawm tsoomfwv hauv biotechnology thiab kev kho mob meej, muaj kev tshawb pom tshiab hauv kev kho cov noob caj noob ces rau ntau yam kab mob, suav nrog cov kab mob caj ces, mob qog noj ntshav, thiab Cov Kab Mob Autoimmune.