A universal plasmid library encoding all permutations of small interfering RNA

Meihong Chen, Lishu Zhang, Hong Yan Zhang, Xiahui Xiong, Bo Wang, Quan Du, Bing Lu, Claes Wahlestedt, Zicai Liang

Research output: Contribution to journalArticlepeer-review

39 Scopus citations

Abstract

Small interfering RNA (siRNA) is normally designed to silence preselected known genes. Such selections are inevitably prone to bias as a result of limited knowledge about the biological process, transcript identity, and functions. A library that contains all permutations of siRNA could avoid such problems. In this paper, it is shown that 5 × 107 siRNA-encoding plasmids can be constructed in a single tube by using vectors with two mutated RNA polymerase III promoters arranged in a convergent manner. Such a library was used to carry out genomewide screening of functional genes in a phenotype-driven manner. Multiple siRNAs that induce a significant increase of cell proliferation speed were identified.

Original languageEnglish (US)
Pages (from-to)2356-2361
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume102
Issue number7
DOIs
StatePublished - Feb 15 2005
Externally publishedYes

Keywords

  • Dual promoters
  • Random targeting
  • Screening
  • Vector

ASJC Scopus subject areas

  • Genetics
  • General

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