The effect of RNA interference induced by inhibition of hTERT and Bi-1 gene expression
10.3969/j.issn.1671-8348.2015.08.002
- VernacularTitle:利用 RNAi阻抑 hTERT 和Bi-1双基因表达的RNAi作用效果的研究
- Author:
Huimin WANG
;
Kexin HE
;
Jianhua XU
;
Chenyu SHANG
;
Keyuan ZHOU
- Publication Type:Journal Article
- Keywords:
hTERT;
bax inhibitor-1;
RNAi
- From:
Chongqing Medicine
2015;(8):1012-1016
- CountryChina
- Language:Chinese
-
Abstract:
Objective In this study ,we constructed a series of recombinant plasmids carriers expressing shRNA targeting hTERT and Bi‐1 gene .These recombinant plasmids carriers were transfected into CNE‐2Z cell lines using Lip and continuously in‐duced the expression of shRNAs .Furthermore ,the shRNAs caused the degradation of mRNAs homologous in sequence with the target genes ,which lead to a sequence‐specific gene silencing .Methods The CNE‐2Z cells was divided into untreated group ,pEG‐FP‐N1 group and pEGFP‐N1/Lip group .Flow cytometry(FCM ) was applied to determine the transfection efficiency .The changes of hTERT and Bi‐1 gene expression were detected by Real‐time RT‐PCR and Western blotting .Results The best transfection effi‐ciency between plasmid and Lip was 2 .5 μg plasmid and 6 .25μL Lip .Conclusion We constructed several shRNA recombinant eu‐karyotic expression plasmids successfully .The recombinant plasmid can inhibit the expression of hTERT and Bi‐1 gene specifically and effectively .
