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Updated: Jun 2, 2026
DNA Vector-based RNA Interference to Study Gene Function in Cancer
Published on: June 4, 2012
[Construction of Cyr61 siRNA expression vector and its characteristic investigation]
Yan Zhang1, Qing-ping Wu, Shang-long Yao
1Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China. yanzizhang917@126.com
Aim:
To construct siRNA recombinant expression vector targeting Cyr61 gene, and to screen the stably transfected cell clone.
Methods:
Three siRNAs sequences based on the sequence of Cyr61mRNA in the GenBank were designed and synthesized, and one scrambled siRNA sequences served as negative control. The cDNA was synthesized and inserted into plasmid pGenesil1.1. The plasmids was sequenced to confirm the inserted sequence.RT-PCR and Western blot analysis was used to assess the levels of Cyr61 mRNA and proteins after the constructed plasmids have been transfected into A549 cells.
Results:
It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting Cyr61 gene was constructed successfully. Inhibition ratio of Cyr61 siRNA at mRNA and protein levels were 59.2% and 52.97%.
Conclusion:
The siRNA recombinant expression vector targeting Cyr61 gene has been constructed successfully and can inhibite the expression of Cyr61 gene in A549 cells significantly.
