[Construction of an expression vector for high expression of siRNA]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Dec;23(12):1117-8.
[Article in Chinese]

Abstract

Aim: To construct the vector for efficient expression of siRNA using pre-mir30 backbone.

Methods: By chemical synthesis method, pre-mir30 backbone introduced an appropriate restriction enzyme site for foreign shRNA inserting was cloned into an expressing vector containing U6 promoter. The silencing efficiency of a new siRNA expressing vector was detected by transfection and Western blot.

Results: The new vector containing pre-mir30 backbone expressing siRNA against GFP could markedly inhibit the expression of GFP compared with the vector expressing control siRNA.

Conclusion: siRNA expressing vector constructed by pre-mir30 backbone could highly express foreign siRNA.

Publication types

  • English Abstract

MeSH terms

  • Animals
  • Cattle
  • DNA Restriction Enzymes / metabolism
  • Gene Expression
  • Genetic Vectors / genetics*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Humans
  • Mice
  • MicroRNAs / genetics*
  • RNA Interference
  • RNA, Small Interfering / biosynthesis*
  • RNA, Small Interfering / genetics*

Substances

  • MicroRNAs
  • RNA, Small Interfering
  • Green Fluorescent Proteins
  • DNA Restriction Enzymes