A universal plasmid library encoding all permutations of small interfering RNA.
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.
Chen
Meihong
M
Chinese Human Genome Center North Beijing, Beijing 100176, People's Republic of China.
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
D001483:Base Sequence; D002460:Cell Line; D049109:Cell Proliferation; D004258:DNA Polymerase III / Q000235:genetics; D015723:Gene Library; D020868:Gene Silencing; D005822:Genetic Vectors; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*; D011401:Promoter Regions, Genetic; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection
D034741:RNA, Small Interfering; D004258:DNA Polymerase III