shRNA vs siRNA
Thaum tus txheej txheem ntawm RNA cuam tshuam (RNAi), kev qhia ntawm lub hom phiaj noob tau poob qis nrog qhov tshwj xeeb thiab kev xaiv. RNAi yog txheej txheem ntuj tsim, thiab nws cuam tshuam me me RNA (siRNA) thiab luv hairpin RNA (shRNA) thiab ob-functional shRNA. Tam sim no, RNAi tau siv dav los ua cov cuab yeej rau kev kho mob qog noj ntshav ntawm tus kheej. Cov ntawv thov ntawm RNAi yog qhov pib ua tiav nrog cov tshuaj sib xyaw ua ke ob-stranded siRNA thiab vector raws li shRNA molecules. Txawm hais tias ob lub molecules no muaj cov txiaj ntsig zoo sib xws, lawv txawv ntawm lawv cov qauv; Yog li, cov txheej txheem molecular ntawm kev txiav txim, RNA txoj hauv kev, thiab cov txiaj ntsig tawm ntawm ob lub molecules kuj txawv.
shRNA
shRNA yog ib ntu ntawm RNA molecule me me uas ua rau cov plaub hau nruj tig uas tuaj yeem siv los ua kom ntsiag to lub hom phiaj gene qhia thaum RNAi. Kev nthuav qhia ntawm shRNA yog ua tiav los ntawm vector, uas tuaj yeem yog tus kab mob lossis kab mob lossis los ntawm kev xa cov plasmids. Lawv yog synthesized nyob rau hauv lub nucleus ntawm hlwb thiab thauj mus rau lub cytoplasm rau cov txheej txheem ntxiv. Cov molecules no muaj txoj hauv kev zoo sib xws ntawm miRNA; Yog li cov synthesis ntawm miRNA tau muab lub hauv paus rau kev nkag siab ntawm shRNA synthesis. RNA polymerase II lossis III tuaj yeem sau cov shRNA los ntawm RNA polymerase II lossis III txhawb nqa. Qhov zoo ntawm kev siv shRNAs yog tias lawv muaj tus nqi qis ntawm degradation thiab turnover. Qhov tsis zoo yog tias nws xav tau kev qhia vector, uas yuav ua rau qee qhov teeb meem kev nyab xeeb.
siRNA
siRNAs yog ob npaug RNA molecules uas muaj li ntawm 20- 25 lub hauv paus khub ntev. Cov no yog siv rau kev tua cov noob los ntawm kev ntsiag to cov noob nrog cov nucleotide sib xyaw ua ke hauv RNAi txoj hauv kev. Gene knockdown los ntawm transfection ntawm siRNA feem ntau ua tsis tiav vim cov nyhuv hloov pauv; tshwj xeeb tshaj yog nyob rau hauv ceev faib cov hlwb thiab cov suppression tsis kav ntev. Txhawm rau kov yeej qhov teeb meem no, siRNA tau hloov kho los ntawm kev nthuav qhia luv luv hairpin qauv. Qhov no hloov molecule ces hu ua shRNA. shRNA yuav tsum tau hloov mus rau siRNA los ntawm Dicer txhawm rau ua nws txoj haujlwm li qub.
Qhov txawv ntawm shRNA thiab siRNA yog dab tsi?
• Tsis zoo li siRNA, shRNA muaj cov qauv hairpin ntxiv. shRNA yog hloov kho version ntawm siRNA.
• shRNA yuav tsum muaj kev qhia vector, whereas siRNA tsis.
• shRNA siv tau rau lub sijhawm ntev knockdown thaum siRNA tsuas yog siv rau lub sijhawm luv luv ntawm cov noob.
• Tsis zoo li cov noob txwv ntawm siRNA, kev tawm tsam ntawm shRNA yog qhov ntev, thiab yog tias nws tau ntxig los ntawm cov kab mob uas tsim nyog, nws tuaj yeem tsim cov tshuaj tua kab mob mus tas li.
• Dicer yuav tsum tau hloov lub shRNA rov qab mus rau siRNA molecule thiaj li ua tiav nws cov haujlwm ib txwm muaj.
