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Updated: Jun 17, 2026
MISSION esiRNA for RNAi Screening in Mammalian Cells
Published on: May 12, 2010
[Construction and identification of eukaryotic expression plasmid expressing siRNA targeting Rac1 and detection of
Mei-Xia Zhang1, Jing Wu, Mei Xin
1Department of Ophthalmology, West China Hospital, Sichuan University, Chengdu 610041, China.
Objective:
To constructed eukaryotic expressing siRNA sections targeting human and rat Rac1 and to observe their effects on Rac1 gene expression in HeLa cell line.
Methods:
siRNA sequences were designed and synthesized targeting human and rat's Rac1 mRNA, and then directionally cloned into pSUPER plasmid. The recombinant vectors were confirmed by enzyme digestion analysis and DNA sequencing. The recombinant vectors were transfected into HeLa cell line and the transfection rate was evaluated with fluorescence microscope. The effects of the recombinants on Rac1 at mRNA levels were observed by RT-PCR.
Results:
Three siRNA expressing recombinants and corresponding negative control vector were constructed and transfected into HeLa cell successfully. Rac1 transcript was reduced significantly in three transfectants.
Conclusion:
The construction of eukaryotic expression vectors expressing siRNA sections targeting Rac1 and identification successfully. This will be very helpful for further study on the function of Rac1 and it may be a useful tool in the treatment of the neovascular retinopathy.
