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2009•Journal of Gannan Medical UniversityRequires access

Construction of the shRNA expression vectors targeting survivin

Fuhua Xie

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Abstract

Objective:To construct the short hairpin RNA(shRNA) expression vector targeting survivin then identify it,which will make the foundation of exploring RNAi-based gene therapy for breast cancer.Methods: Eight strands of DNA fragement with the structure of survivin shRNA were designed and synthesized.After annealed,DNA strands were cloned into the pSilencer adeno 1.0-CMV system,then transformed DH5α with the recombinants.The plasmids were amplified and extracted so as to identify by electrophoresis and sequencement.Results: We found the expression vectors were constructed successfully.Conclusion: The construction of the vector paved a way for the further research,in which the recombinants will be packed with adenovirus and infect nude mice bearing human breast cancer.

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What this paper is about

Objective:To construct the short hairpin RNA(shRNA) expression vector targeting survivin then identify it,which will make the foundation of exploring RNAi-based gene therapy for breast cancer.Methods: Eight strands of DNA fragement with the structure of survivin shRNA were designed and synthesized.After annealed,DNA strands were cloned into the pSilencer adeno 1.0-CMV system,then transformed DH5α with the recombinants.The plasmids were amplified and extracted so as to identify by electrophoresis and sequencement.Results: We found the expression vectors were constructed successfully.Conclusion: The construction of the vector paved a way for the further research,in which the recombinants will be packed with adenovirus and infect nude mice bearing human breast cancer.

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Available abstract

Objective:To construct the short hairpin RNA(shRNA) expression vector targeting survivin then identify it,which will make the foundation of exploring RNAi-based gene therapy for breast cancer.Methods: Eight strands of DNA fragement with the structure of survivin shRNA were designed and synthesized.After annealed,DNA strands were cloned into the pSilencer adeno 1.0-CMV system,then transformed DH5α with the recombinants.The plasmids were amplified and extracted so as to identify by electrophoresis and sequencement.Results: We found the expression vectors were constructed successfully.Conclusion: The construction of the vector paved a way for the further research,in which the recombinants will be packed with adenovirus and infect nude mice bearing human breast cancer.

Key concepts: Small hairpin RNA, Survivin, Vector (molecular biology), Genetic enhancement, RNA interference, Medicine, DNA, Viral vector

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