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- Title
Second-generation shRNA libraries covering the mouse and human genomes.
- Authors
Silva, Jose M.; Li, Mamie Z.; Chang, Ken; Wei Ge; Golding, Michael C.; Rickles, Richard J.; Siolas, Despina; Hu, Guang; Paddison, Patrick J; Schlabach, Michael R; Sheth, Nihar; Bradshaw, Jeff; Burchard, Julia; Kulkarni, Amit; Cavet, Guy; Sachidanandam, Ravi; McCombie, W. Richard; Cleary, Michele A.; Elledge, Stephen J.; Hannon, Gregory J.
- Abstract
Loss-of-function phenotypes often hold the key to understanding the connections and biological functions of biochemical pathways. We and others previously constructed libraries of short hairpin RNAs that allow systematic analysis of RNA interference–induced phenotypes in mammalian cells. Here we report the construction and validation of second-generation short hairpin RNA expression libraries designed using an increased knowledge of RNA interference biochemistry. These constructs include silencing triggers designed to mimic a natural microRNA primary transcript, and each target sequence was selected on the basis of thermodynamic criteria for optimal small RNA performance. Biochemical and phenotypic assays indicate that the new libraries are substantially improved over first-generation reagents. We generated large-scale-arrayed, sequence-verified libraries comprising more than 140,000 second-generation short hairpin RNA expression plasmids, covering a substantial fraction of all predicted genes in the human and mouse genomes. These libraries are available to the scientific community.
- Subjects
PHENOTYPES; HUMAN genome; GENE expression; TRANSCRIPTION factors; ANIMAL models in research; GENETICS
- Publication
Nature Genetics, 2005, Vol 37, Issue 11, p1281
- ISSN
1061-4036
- Publication type
Article
- DOI
10.1038/ng1650