Design and cloning of lentiviral vectors expressing small interfering RNAs.

by: G Tiscornia, O Singer, IM Verma

Nat Protoc, Vol. 1, No. 1. (2006), pp. 234-240.

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Abstract

RNA interference (RNAi) has emerged as a powerful technique to downregulate gene expression. The use of polIII promoters to express small hairpin RNAs (shRNAs), combined with the versatility and robustness of lentiviral vector-mediated gene delivery to a wide range of cell types offers the possibility of long-term downregulation of specific target genes both in vitro and in vivo. The use of silencing lentivectors allows for a rapid and convenient way of establishing cell lines (or transgenic mice) that stably express shRNAs for analysis of phenotypes produced by knockdown of a gene product. Here we present two possible protocols describing the design and cloning of silencing lentiviral vectors. These protocols can be completed in less than 3 weeks.

@article{citeulike:1675961, abstract = {RNA interference (RNAi) has emerged as a powerful technique to downregulate gene expression. The use of polIII promoters to express small hairpin RNAs (shRNAs), combined with the versatility and robustness of lentiviral vector-mediated gene delivery to a wide range of cell types offers the possibility of long-term downregulation of specific target genes both in vitro and in vivo. The use of silencing lentivectors allows for a rapid and convenient way of establishing cell lines (or transgenic mice) that stably express shRNAs for analysis of phenotypes produced by knockdown of a gene product. Here we present two possible protocols describing the design and cloning of silencing lentiviral vectors. These protocols can be completed in less than 3 weeks.}, address = {The Salk Institute for Biological Studies, Laboratory of Genetics, 10010 North Torrey Pines Road, La Jolla, California 92037, USA.}, author = {Tiscornia, G. and Singer, O. and Verma, I. M. }, citeulike-article-id = {1675961}, doi = {http://dx.doi.org/10.1038/nprot.2006.36}, issn = {1750-2799}, journal = {Nat Protoc}, keywords = {cloning, lentivirus, rnai, shrna}, number = {1}, pages = {234--240}, posted-at = {2008-02-24 02:19:25}, priority = {2}, title = {Design and cloning of lentiviral vectors expressing small interfering RNAs.}, url = {http://dx.doi.org/10.1038/nprot.2006.36}, volume = {1}, year = {2006} }

RIS record

TY - JOUR ID - citeulike:1675961 L3 - citeulike-article-id:1675961 TI - Design and cloning of lentiviral vectors expressing small interfering RNAs. JF - Nat Protoc VL - 1 IS - 1 SP - 234 EP - 240 AD - The Salk Institute for Biological Studies, Laboratory of Genetics, 10010 North Torrey Pines Road, La Jolla, California 92037, USA. SN - 1750-2799 N2 - RNA interference (RNAi) has emerged as a powerful technique to downregulate gene expression. The use of polIII promoters to express small hairpin RNAs (shRNAs), combined with the versatility and robustness of lentiviral vector-mediated gene delivery to a wide range of cell types offers the possibility of long-term downregulation of specific target genes both in vitro and in vivo. The use of silencing lentivectors allows for a rapid and convenient way of establishing cell lines (or transgenic mice) that stably express shRNAs for analysis of phenotypes produced by knockdown of a gene product. Here we present two possible protocols describing the design and cloning of silencing lentiviral vectors. These protocols can be completed in less than 3 weeks. KW - cloning KW - lentivirus KW - rnai KW - shrna AU - Tiscornia, G AU - Singer, O AU - Verma, IM PY - 2006/// UR - http://dx.doi.org/10.1038/nprot.2006.36 ER -

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