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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 and George Klein
Proceedings of the National Academy of Sciences of the United States of America
Vol. 102, No. 7 (Feb. 15, 2005), pp. 2356-2361
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/3374619
Page Count: 6
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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.
Proceedings of the National Academy of Sciences of the United States of America © 2005 National Academy of Sciences