Restriction enzyme–generated siRNA (REGS) vectors and libraries View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2004-02

AUTHORS

George Sen, Tom S Wehrman, Jason W Myers, Helen M Blau

ABSTRACT

Small interfering RNA (siRNA) technology facilitates the study of loss of gene function in mammalian cells and animal models, but generating multiple siRNA vectors using oligonucleotides is slow, inefficient and costly. Here we describe a new, enzyme-mediated method for generating numerous functional siRNA constructs from any gene of interest or pool of genes. To test our restriction enzyme-generated siRNA (REGS) system, we silenced a transgene and two endogenous genes and obtained the predicted phenotypes. REGS generated on average 34 unique siRNAs per kilobase of sequence. REGS enabled us to create enzymatically a complex siRNA library (>4 x 10(5) clones) from double-stranded cDNA encompassing known and unknown genes with 96% of the clones containing inserts of the appropriate size. More... »

PAGES

183-189

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/ng1288

DOI

http://dx.doi.org/10.1038/ng1288

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1014456336

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/14704668


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42 schema:description Small interfering RNA (siRNA) technology facilitates the study of loss of gene function in mammalian cells and animal models, but generating multiple siRNA vectors using oligonucleotides is slow, inefficient and costly. Here we describe a new, enzyme-mediated method for generating numerous functional siRNA constructs from any gene of interest or pool of genes. To test our restriction enzyme-generated siRNA (REGS) system, we silenced a transgene and two endogenous genes and obtained the predicted phenotypes. REGS generated on average 34 unique siRNAs per kilobase of sequence. REGS enabled us to create enzymatically a complex siRNA library (>4 x 10(5) clones) from double-stranded cDNA encompassing known and unknown genes with 96% of the clones containing inserts of the appropriate size.
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