Construction and Screening of Recombinant Interference Vector of Pluripotency Associated Genes cNanog and cPouV in Chicken Embryonic Stem Cells

Abstract

Abstract: The study was to construct the siRNA(small interfering RNA)expression vectors targeting cNanog and cPouV of Chicken Embryonic Stem cells(CES cells), and evaluate its inhibitory effect on cNanog and cPouV in 293T cells. Three regions of cNanog and cPouV gene were respectively selected to synthesize oligonucleotides, and construct interference vectors. These vectors were respectively transfected into 293T cells as whole-gene expression vectors were transduced simultaneously. Western blotting analysis showed target genes expression were downregulated after RNAi and the oligonucleotide sequence S3 was found to be the most efficient one in interfering effect. The results of this experiment indicate siRNA or shRNA(short hairping RNA)were translated into cells and suppress the expression of target genes.

Key words: Chicken Embryonic, Stem, Cells, CNanog, CPouV Interference vector

Peng Te, Wang Bingyun, Li Dongsheng, Chen Zhisheng, Chen Shengfeng, Ji Huiqin, Chen Jinding. Construction and Screening of Recombinant Interference Vector of Pluripotency Associated Genes cNanog and cPouV in Chicken Embryonic Stem Cells[J]. Biotechnology Bulletin, 2014, 0(5): 117-121.

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