Qhov sib txawv tseem ceeb ntawm ZFN TALEN thiab CRISPR yog tias ZFN yog tib neeg tsim cov noob kho cov txheej txheem raws li Zinc ntiv tes nucleases tsim los ntawm zinc ntiv tes sau thiab Fok1 endonuclease domain. Meanwhile, TALEN yog ib tug txiv neej tsim noob kho cov txheej txheem raws li fusion proteins uas muaj cov kab mob TALE protein thiab Fok1 endonuclease, thiab CRISPR yog lub ntuj RNA raws li kab mob tiv thaiv mechanism uas yog tsav los ntawm ob hom RNAs thiab txuam nrog Cas proteins.
Gene kho cov cuab yeej yog ua raws li cov txheej txheem tsim nucleases uas muaj cov kab ke tshwj xeeb khi cov thawj thiab cov tsis tshwj xeeb DNA cleavage modules. ZFN, TALEN, thiab CRISPR yog peb hom kev kho cov noob caj noob ces. ZEN thiab TALEN yog cov txheej txheem tsim los ntawm tib neeg, thaum CRISPR yog ib qho kev tshwm sim hauv cov kab mob. Tag nrho peb lub tshuab lees paub cov kab ke thiab tshem tawm lub hom phiaj DNA. Zinc ntiv tes nucleases yog tsim los ntawm zinc ntiv tes sau thiab Fok1 endonuclease. TALENs yog tsim los ntawm cov kab mob TALE protein thiab Fok1 endonuclease. CRISPR yog tsim los ntawm ob RNAs (trans-activating crRNA thiab ib qho kev qhia RNA).
ZFN yog dab tsi?
ZFN yog cov cuab yeej kho cov noob raws li zinc-ntiv tes nucleases. Zinc ntiv tes nucleases yog chimeric nucleases tsim los ntawm cov kab ke tshwj xeeb DNA-binding domains thiab non-specific DNA cleaving domains los ntawm FokI txwv endonuclease. Qhov no yog tib neeg tsim genome kho tshuab uas ua rau muaj ntau yam kev hloov pauv noob los ntawm kev ua kom ob txoj hlua tawg hauv DNA.
Daim duab 01: ZFN
ZFN yog ib qho cuab yeej tseem ceeb thaum qhia txog kev hloov pauv hauv cov nroj tsuag thiab ntau hom qoob loo. Nws tso cai rau kev koom ua ke ntawm cov txiaj ntsig zoo rau hauv cov nroj tsuag. Ntxiv mus, ZFN muaj peev xwm kho tau vim nws muaj peev xwm ua tau qhov hloov kho caj ces kom tshem tawm cov tsos mob zoo.
TALEN yog dab tsi?
Kev sau npe ua kom zoo li nucleases (TALEN) yog cov cuab yeej kho cov noob uas yog raws li chimeric nucleases zoo ib yam li ZFN. Nws tseem yog tib neeg tsim genome kho tshuab. TALEN induces gene modification los ntawm inducing ob-strand so nyob rau hauv DNA. TALENs yog tsim los ntawm cov kab ke tshwj xeeb DNA-binding domains fused mus rau qhov tsis yog DNA cleavage module. Yog li ntawd, nws muaj FokI cleavage domains thiab DNA-binding domains muab los ntawm TALE proteins.
Daim duab 02: TALEN
TALE proteins yog ib txwm tshwm sim hauv cov kab mob genus Xanthomonas. Cov proteins no muaj DNA binding domains uas muaj ib tug series ntawm 33-35 amino acid rov domains. Ib qho txiaj ntsig tseem ceeb ntawm TALEN dhau ZFN yog qhov kev sib khi tshwj xeeb ntawm DNA khi cov npe tuaj yeem hloov kho hauv TALEN los ntawm kev hloov kho rau 33-34 amino acid rov ua dua.
CRISPR yog dab tsi?
CRISPR sawv cev rau Clustered Regulatory Interspaced Short Palindromic Repeats. Nws yog lub ntuj RNA raws li kev tiv thaiv mechanism ua rau cov kab mob. Qhov tseeb, nws yog genome kho tshuab siv tshwj xeeb tshaj yog tawm tsam cov kab mob. Tus kab mob los yog tus neeg tawm tsam txawv teb chaws tuaj yeem qhib qhov system no hauv cov kab mob. CRISPR ua haujlwm ua ke nrog CRISPR-associated protein (Cas9).
CRISPR muaj ob RNAs: trans-activating crRNA thiab ib qho kev qhia RNA. Yog li, kev lees paub ntawm CRISPR yog nyob ntawm ob qho ntawm RNAs. Thaum cov kab mob DNA nkag mus rau hauv cov kab mob ntawm tes, CRISPR locus tsim cov lus qhia RNA, uas escorts Cas cov proteins kom ncav cuag thiab khi nrog cov kab mob DNA thiab cleave ntawm ib qho chaw tshwj xeeb. Thaum cov kab mob DNA tau cleaved, nws yuav tsis ua haujlwm vim yog kev ntsiag to ntawm cov kab mob DNA. Li no, CRISPR system ntes, cleaves thiab inactivates kab mob nrog kev pab los ntawm Cas proteins.
Daim duab 03: CRISPR/Cas9
CRISPR/Cas9 yog siv los ua genome kho qhov system. Nws yog kev siv tshuab tsis ntev los no. Cov thev naus laus zis no yooj yim heev, ua haujlwm tau zoo, thiab siv nyiaj ntau dua lwm cov txheej txheem kho genome. Nyob rau hauv kev sib piv rau ZFN thiab TALEN, CRISPR / Cas9 system yog ib qho cuab yeej siv tau zoo hauv kev tsim cov kev hloov pauv ntawm cov genome.
Qhov Zoo sib xws ntawm ZFN TALEN thiab CRISPR yog dab tsi?
- ZFN, TALEN, thiab CRISPR yog cov txheej txheem kho cov noob.
- Tag nrho peb cov cuab yeej yuav muaj kev cog lus hauv kev kho cov caj ces ua rau muaj ntau yam kab mob.
- Txawm li cas los xij, lawv qhia txog kev txwv hauv kev tsom, qhov tsis zoo, thiab cov hom phiaj.
- Txhua peb lub tshuab tuaj yeem ua rau ob txoj hlua tawg.
Qhov txawv ntawm ZFN TALEN thiab CRISPR yog dab tsi?
ZFN yog cov txheej txheem kho cov noob raws li Zinc ntiv tes nucleases thaum TALEN yog cov txheej txheem kho noob raws li fusion proteins uas muaj cov kab mob TALE protein thiab Fok1 endonuclease, thiab CRISPR yog lub ntuj RNA raws li cov kab mob tiv thaiv kab mob uas tau tsav. los ntawm ob hom RNA thiab txuam nrog Cas proteins. Yog li, qhov no yog qhov sib txawv tseem ceeb ntawm ZFN TALEN thiab CRISPR.
Cov hauv qab no infographic teev qhov sib txawv ntawm ZFN TALEN thiab CRISPR hauv daim ntawv teev npe rau ib sab ntawm kev sib piv.
Summary – ZFN vs TALEN vs CRISPR
ZFN, TALEN, thiab CRISPR yog gene kho thev naus laus zis. CRISPR yog cov thev naus laus zis tsis ntev los no uas qhia tau tias muaj txiaj ntsig zoo dua, muaj peev xwm ua tau, thiab kev siv ntau lub luag haujlwm. ZFN thiab TALEN yog tib neeg tsim cov cuab yeej cuab tam, thaum CRISPR yog cov kab mob tiv thaiv kab mob. Ob leeg ZFN thiab TALEN yog engineered nucleases. CRISPR muaj ob hom RNA txuam nrog Cas proteins. Yog li, qhov no yog cov ntsiab lus ntawm qhov sib txawv ntawm ZFN TALEN thiab CRISPR.
