A CPLs vector-based RNAi technology to inhibit the expression of rat CⅡTA and MHCⅡ genes
10.3760/cma.j.issn.0254-5101.2014.08.005
- VernacularTitle:阳离子高分子脂质体介导的 RNA 干扰技术对大鼠 CⅡTA 和 MHCⅡ基因表达的抑制作用
- Author:
Jianlin WANG
;
Gang LIU
;
Chengmei ZHAO
;
Yujie QIU
;
Liwei ZHU
- Publication Type:Journal Article
- Keywords:
RNA interference;
CⅡTA;
MHCⅡ;
Cationic polymeric liposomes
- From:
Chinese Journal of Microbiology and Immunology
2014;(8):594-598
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effects of cationic polymeric liposomes (CPLs) vector-based RNA interference (RNAi) technology on the expression of rat MHCⅡ transactivator ( CⅡTA) and MHCⅡgenes .Methods According to the genetic information of CⅡTA downloaded from GenBank, three short hairpin RNA (shRNA) sequences targeting CⅡTA sequences were designed .CPLs vectors were constructed and coupled to shRNA plasmid vectors to form pCⅡTA-CPLs vectors .Six groups including control group , CPLs control group , pHK-CⅡTA control group and three pCⅡTA-CPLs groups were set up.Rat dendritic cells (DCs) were transfected in vitro.Real time PCR and flow cytometry analysis were used to detect the expression of CⅡTA and MHCⅡat mRNA and protein levels in DCs after transfec-tion.Results The pCⅡTA-CPLs vectors were successfully constructed .Compared with control groups ,the transcription level of CⅡTA and MHCⅡand the expression of MHCⅡat protein level were significantly in-hibited in all pCⅡ TA-CPLs groups ( P<0 .01 ) .The strongest inhibitory effects of pCⅡTA-CPLs on the ex-pression of CⅡTA and MHCⅡgenes were observed in the second pCⅡTA-CPLs group.There was a positive correlation between the expression of CⅡTA and MHCⅡ.Conclusion CPLs vectors were effective gene carriers.The constructed pCⅡTA-CPLs vectors significantly inhibited the in vitro expression of rat CⅡTA and MHCⅡ, which provided evidences for further investigation on pCPLs-CⅡTA vectors in vivo.
