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

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 × 10 7 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

  • Base Sequence
  • Cell Line
  • Cell Proliferation
  • DNA Polymerase III/genetics
  • Gene Library
  • Gene Silencing
  • Genetic Vectors
  • Humans
  • Molecular Sequence Data
  • Plasmids/genetics
  • Promoter Regions, Genetic
  • RNA, Small Interfering/genetics
  • Transfection
  • Screening
  • Random targeting
  • Dual promoters
  • Vector

ASJC Scopus subject areas

  • General

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