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2004•Unpublished venueRequires access

A method of siRNA synthesis by T7 RNA polymerase transcription

Peng Jian

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Abstract

Objective: To establish a method using T7 RNA polymerase for obtaining large amount of siRNA. Method: The siRNA(small interfering RNA) targeting luciferase was synthesized by T7 RNA polymerase transcription in vitro. The siRNA was transfected into HepG2 cells via lipofectamine method. The inhibitory effects of siRNA were evaluated by measuring luciferase light intensity. Result: The siRNA displayed inhibitory effects on the luciferase expression with the inhibitory rate of 80%. Conclusion: A method of siRNA synthesis in vitro was established that affords large amounts of double-stranded RNA cheaply and conveniently for RNA interference.

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What this paper is about

Objective: To establish a method using T7 RNA polymerase for obtaining large amount of siRNA. Method: The siRNA(small interfering RNA) targeting luciferase was synthesized by T7 RNA polymerase transcription in vitro. The siRNA was transfected into HepG2 cells via lipofectamine method. The inhibitory effects of siRNA were evaluated by measuring luciferase light intensity. Result: The siRNA displayed inhibitory effects on the luciferase expression with the inhibitory rate of 80%. Conclusion: A method of siRNA synthesis in vitro was established that affords large amounts of double-stranded RNA cheaply and conveniently for RNA interference.

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Available abstract

Objective: To establish a method using T7 RNA polymerase for obtaining large amount of siRNA. Method: The siRNA(small interfering RNA) targeting luciferase was synthesized by T7 RNA polymerase transcription in vitro. The siRNA was transfected into HepG2 cells via lipofectamine method. The inhibitory effects of siRNA were evaluated by measuring luciferase light intensity. Result: The siRNA displayed inhibitory effects on the luciferase expression with the inhibitory rate of 80%. Conclusion: A method of siRNA synthesis in vitro was established that affords large amounts of double-stranded RNA cheaply and conveniently for RNA interference.

Key concepts: Lipofectamine, T7 RNA polymerase, Small interfering RNA, Luciferase, Molecular biology, RNA interference, Transcription (linguistics), RNA

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