[Recombinant adenovirus mediate RNA silencing of perforin in NK-92 cells and its function].
OBJECTIVE
To adenovirus-mediated RNAi technology be used to down-regulate perforin expression in NK-92 cells. This study will provide convenience for further investigation of the mechanism of perforin.


METHODS
First, we designed three perforin shRNAs (shPF) and identified their knockdown efficiency at mRNA levels. Then we constructed recombinant adenovirus with shPF. After that, we transfected NK-92 cells with Ad-shPF and detected the expression of perforin at mRNA and protein levels.


RESULTS
We confirm that the design of shPF sequences are effective. Moreover, the mRNA level of NK-92 cells treated by Ad-shPFP1 drop to 32% compare to Ad-scramble treated control. Furthermore, using Western blot, we find Ad-shPF1 treatment can obviously down-regulate the expression of perforin protein.


CONCLUSIONS
Recombinant adenovirus with perforin shRNA can knockdown the expression of perforin in NK-92 cells.
Zhao
Li-lin
LL
First Affiliated Hospital, Chongqing Medical University, Chongqing, China. zhaolilin263@163.com
Liu
Yang
Y
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism; D054353:Perforin / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D014161:Transduction, Genetic / Q000379:methods*
D054353:Perforin