1. A universal plasmid library encoding all permutations of small interfering RNA
- Author
-
Bo Wang, Xiahui Xiong, Quan Du, Claes Wahlestedt, Bing Lu, Hong-Yan Zhang, Zicai Liang, Meihong Chen, and Lishu Zhang
- Subjects
Genetics ,Small interfering RNA ,Multidisciplinary ,Base Sequence ,Trans-acting siRNA ,Genetic Vectors ,Molecular Sequence Data ,RNA ,Biology ,Biological Sciences ,Transfection ,RNA polymerase III ,Cell Line ,RNA silencing ,Plasmid ,Humans ,Genomic library ,Gene Silencing ,RNA, Small Interfering ,Promoter Regions, Genetic ,Gene ,Cell Proliferation ,DNA Polymerase III ,Gene Library ,Plasmids - 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.
- Published
- 2005