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Comparison of bovine RNA polymerase III promoters for short hairpin RNA expression

journal contribution
posted on 2006-08-01, 00:00 authored by L Lambeth, T Wise, R Moore, Morley Muralitharan, T Doran
RNA interference (RNAi) mediated by DNA-based expression of short hairpin RNA (shRNA) is a powerful method of sequence-specific gene knockdown. A number of vectors for expression of shRNA have been developed that feature promoters from RNA polymerase III (pol III)-transcribed genes of mouse or human origin. To advance the use of RNAi as a tool for functional genomic research and for future development of specific therapeutics in the bovine species, we have developed shRNA expression vectors that feature novel bovine RNA pol III promoters. We characterized two bovine U6 small nuclear RNA (snRNA) promoters (bU6-2 and bU6-3) and a bovine 7SK snRNA promoter (b7SK). We compared the efficiency of each of these promoters to express shRNA molecules. Promoter activity was measured in the context of RNAi by targeting and suppressing the reporter gene encoding enhanced green fluorescent protein. Results show that the b7SK promoter induced the greatest level of suppression in a range of cell lines. The comparison of these bovine promoters in shRNA expression is an important component for the future development of bovine-specific RNAi-based research.

History

Journal

Animal genetics

Volume

37

Issue

4

Pagination

369 - 372

Publisher

Blackwell Publishing

Location

Oxford, U.K.

ISSN

0268-9146

eISSN

1365-2052

Language

eng

Publication classification

C1 Refereed article in a scholarly journal

Copyright notice

2006, Blackwell Publishing