Construction and expression of small interfering RNA expression vector of CXCR4
- VernacularTitle:趋化因子受体CXCR4小分子干扰RNA表达载体的构建及表达
- Author:
Yan LIU
;
Long WANG
;
Dawei ZHANG
;
Deming ZHAO
- Publication Type:Journal Article
- Keywords:
receptor,chemokine;
HIV-1;
RNA interference;
small interfering RNA
- From:
Journal of Jilin University(Medicine Edition)
2006;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the recombinant plasmid carrying small interfering RNA(siRNA) to CXCR4 and observe dumbness effect of chemokine receptor gene CXCR4 suppression by RNA interference(RNAi),and explore the new method of more efficacious therapeutic possibilities for HIV-1.Methods Small interfering RNAs targeting CXCR4 gene were designed by using software.The complement form was obtained by annealing and interted into vector Psilencer3.1-H1neo.After sequencing,MT4 cells were transfected using the plasmid vector.RT-PCR and flow cytometry detection were used to evaluate the suppression of CXCX4 expression in different groups.Results The recombinant plasmid had the correct special fragments and DNA sequence detected by PCR,electrophoresis and DNA sequencing;The siRNA obviously suppressed the expression of CXCR4.When transfected with plasmid vector for 48 h,the inhibitory rate was 70%,compared with control and negative groups,there were significant differences(P
