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.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 2356-2361 |
| Number of pages | 6 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 102 |
| Issue number | 7 |
| DOIs | |
| State | Published - Feb 15 2005 |
| Externally published | Yes |
Keywords
- Base Sequence
- Cell Line
- Cell Proliferation
- DNA Polymerase III/genetics
- Gene Library
- Gene Silencing
- Genetic Vectors
- Humans
- Molecular Sequence Data
- Plasmids/genetics
- Promoter Regions, Genetic
- RNA, Small Interfering/genetics
- Transfection
- Screening
- Random targeting
- Dual promoters
- Vector
ASJC Scopus subject areas
- General