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A universal plasmid library encoding all permutations of small interfering RNA.

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 x 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.

SUBMITTER: Chen M

PROVIDER: S-EPMC548965 | biostudies-literature | 2005 Feb

REPOSITORIES: biostudies-literature

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A universal plasmid library encoding all permutations of small interfering RNA.

Chen Meihong M Zhang Lishu L Zhang Hong-Yan HY Xiong Xiahui X Wang Bo B Du Quan Q Lu Bing B Wahlestedt Claes C Liang Zicai Z

Proceedings of the National Academy of Sciences of the United States of America 20050204 7

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 x 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 ma ...[more]

PMID: 15695593

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