[Inhibitory effect of short hairpin RNA on expression of Hax-1 gene in HeLa cells].
OBJECTIVE
To construct the short hairpin RNA (shRNA) eukaryotic expression vector specific for HS1-associated protein X-1 (Hax-1) and investigate the inhibitory effect of it on Hax-1.


METHODS
According to the design rules of shRNA, the specific sequence of 19 nucleotides were selected from Hax-1 cDNA sequence and designed as the cDNA template of siRNA. ShRNA vector pGenesil-Hax-1 was constructed by recombining the synthesized specific sequence with siRNA expression vector pGensil-1. After the recombinant plasmids were transfected into HeLa cells, RT-PCR technique and Western blot were applied to analyze mRNA and protein expression of Hax-1.


RESULTS
The results of RT-PCR showed that the down-regulation of Hax-1 mRNA expression was found in the pGenesil-Hax-1 transfected group, but not in the pGenesil-1 transfected group or the negative control group (P<0.01). The expression of Hax-1 protein decreased by 70% in the pGenesil-Hax-1 transfected group compared with the negative control group.


CONCLUSIONS
Hax-1 gene expression can be inhibited markedly by specific shRNA in HeLa cells, which establishes the experimental foundation for further study on the biological functions of Hax-1 in HeLa cells.
Zhai
Zhi-Fang
ZF
Department of Dermatology, Southwest Hospital, Third Military Medical University, Chongqing, China. zhaizf1004@163.com
Hao
Fei
F
Jiang
You-Zhao
YZ
Zhong
Bai-Yu
BY
Yan
Heng
H
Zhong
Hua
H
D048868:Adaptor Proteins, Signal Transducing; D015536:Down-Regulation; D055785:Gene Knockdown Techniques / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D055029:Inverted Repeat Sequences / Q000235:genetics*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000737:chemistry / Q000235:genetics*; D014162:Transfection
D048868:Adaptor Proteins, Signal Transducing; C104759:HAX1 protein, human; D011506:Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering