Synthetic shRNAs as potent RNAi triggers

Siolas, D., Lerner, C., Burchard, J., Ge, W., Linsley, P. S., Paddison, P. J., Hannon, G. J., Cleary, M. A. (February 2005) Synthetic shRNAs as potent RNAi triggers. Nat Biotechnol, 23 (2). pp. 227-31. ISSN 1087-0156 (Print)

URL: http://www.ncbi.nlm.nih.gov/pubmed/15619616
DOI: 10.1038/nbt1052

Abstract

Designing potent silencing triggers is key to the successful application of RNA interference (RNAi) in mammals. Recent studies suggest that the assembly of RNAi effector complexes is coupled to Dicer cleavage. Here we examine whether transfection of optimized Dicer substrates results in an improved RNAi response. Dicer cleavage of chemically synthesized short hairpin RNAs (shRNAs) with 29-base-pair stems and 2-nucleotide 3' overhangs produced predictable homogeneous small RNAs comprising the 22 bases at the 3' end of the stem. Consequently, direct comparisons of synthetic small interfering RNAs and shRNAs that yield the same small RNA became possible. We found synthetic 29-mer shRNAs to be more potent inducers of RNAi than small interfering RNAs. Maximal inhibition of target genes was achieved at lower concentrations and silencing at 24 h was often greater. These studies provide the basis for an improved approach to triggering experimental silencing via the RNAi pathway.

Item Type: Paper
Additional Information:
Uncontrolled Keywords: Gene Expression Regulation genetics Gene Silencing physiology Gene Targeting methods Genetic Engineering methods RNA Small Interfering chemistry genetics Transfection methods
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNAi
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > dicer
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > dicer

bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > shRNA
CSHL Authors:
Communities: CSHL labs > Hannon lab
Watson School > Publications
Depositing User: CSHL Librarian
Date: February 2005
Date Deposited: 05 Jan 2012 19:26
Last Modified: 19 Sep 2014 15:53
Related URLs:
URI: http://repository.cshl.edu/id/eprint/22704

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