[Universal plasmid library encoding all permutations of siRNA]
>>Published online before print February 4, 2005, 10.1073/pnas.0401549101
PNAS | February 15, 2005 | vol. 102 | no. 7 | 2356-2361
BIOCHEMISTRY
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 ¶ and Zicai Liang *, , ||
*Chinese Human Genome Center North Beijing, Beijing 100176, People's Republic of China; Center for Genomics and Bioinformatics, Karolinska Institutet, 171 77 Stockholm, Sweden; and Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, National Laboratory of Medical Molecular Biology, Beijing 100005, People's Republic of China
Edited by George Klein, Karolinska Institutet, Stockholm, Sweden and approved October 19, 2004 (received for review March 4, 2004)
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 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. <<
This article is identified by PNAS as one of the most widely read in February.
Cheers, Tuck |
