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2007•Journal of Zhengzhou UniversityRequires access

Vector construction and silencing effect of VEGF gene targeted small interfering RNA

Pei-Jie Ding

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Abstract

Aim: To construct VEGF gene-targeted small interfering RNA(siRNA) vector. Methods: VEGF gene-targeted hairpin siRNA was designed, then two complementary oligo nucleotide strand were synthesized and inserted into pSUPERneo-GFP vector after annealing to get the new vector pSUPERneo-GFP-siVEGF, which was digested by restrictive enzyme and sequenced. After that, the new vector was transfected into human esophageal carcinoma Eca109 cell using lipofectamine method, and the mRNA exprssion level of VEGF gene in cells was detected by RT-PCR.Results and Conclusion: pSUPERneo-GFP-siVEGF is built successfully,and the expression level of VEGF mRNA in transfected Eca109 was decreased.

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What this paper is about

Aim: To construct VEGF gene-targeted small interfering RNA(siRNA) vector. Methods: VEGF gene-targeted hairpin siRNA was designed, then two complementary oligo nucleotide strand were synthesized and inserted into pSUPERneo-GFP vector after annealing to get the new vector pSUPERneo-GFP-siVEGF, which was digested by restrictive enzyme and sequenced. After that, the new vector was transfected into human esophageal carcinoma Eca109 cell using lipofectamine method, and the mRNA exprssion level of VEGF gene in cells was detected by RT-PCR.Results and Conclusion: pSUPERneo-GFP-siVEGF is built successfully,and the expression level of VEGF mRNA in transfected Eca109 was decreased.

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Available abstract

Aim: To construct VEGF gene-targeted small interfering RNA(siRNA) vector. Methods: VEGF gene-targeted hairpin siRNA was designed, then two complementary oligo nucleotide strand were synthesized and inserted into pSUPERneo-GFP vector after annealing to get the new vector pSUPERneo-GFP-siVEGF, which was digested by restrictive enzyme and sequenced. After that, the new vector was transfected into human esophageal carcinoma Eca109 cell using lipofectamine method, and the mRNA exprssion level of VEGF gene in cells was detected by RT-PCR.Results and Conclusion: pSUPERneo-GFP-siVEGF is built successfully,and the expression level of VEGF mRNA in transfected Eca109 was decreased.

Key concepts: Lipofectamine, Transfection, Molecular biology, Small interfering RNA, Gene silencing, Messenger RNA, Small hairpin RNA, Gene

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