Strategies for Generation of an siRNA Expression Library Directed Against the Human Genome
- 1 October 2003
- journal article
- research article
- Published by Mary Ann Liebert Inc in Oligonucleotides
- Vol. 13 (5), 325-333
- https://doi.org/10.1089/154545703322617005
Abstract
RNA interference (RNAi) is a phenomenon whereby expression of an individual gene is specifically silenced by the introduction of a double-stranded RNA (dsRNA) whose sequence is homologous to that of the gene in question. The generation of a small interfering RNA (siRNA) expression library directed against the entire human genome is a project that requires solutions to many difficult technical problems. We present here some strategies for solving some of these problems, including the development of genetically stable and highly active siRNA expression vectors, a procedure for selection of favorable target sites, and an efficient and inexpensive procedure for constructing an siRNA expression library.Keywords
This publication has 29 references indexed in Scilit:
- Short hairpin type of dsRNAs that are controlled by tRNAVal promoter significantly induce RNAi-mediated gene silencing in the cytoplasm of human cellsNucleic Acids Research, 2003
- Identification of genes by hybrid ribozymes that couple cleavage activity with the unwinding activity of an endogenous RNA helicaseEMBO Reports, 2002
- A System for Stable Expression of Short Interfering RNAs in Mammalian CellsScience, 2002
- Identification of genes that function in the TNF-α-mediated apoptotic pathway using randomized hybrid ribozyme librariesNature Biotechnology, 2002
- Specific inhibition of gene expression by small double-stranded RNAs in invertebrate and vertebrate systemsProceedings of the National Academy of Sciences, 2001
- Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cellsNature, 2001
- Identification of Id4 as a regulator of BRCA1 expression by using a ribozyme-library-based inverse genomics approachProceedings of the National Academy of Sciences, 2001
- Functional genomic analysis of cell division in C. elegans using RNAi of genes on chromosome IIINature, 2000
- A transcription reinitiation intermediate that is stabilized by activatorNature, 2000
- Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegansNature, 1998