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Characterization of zebrafish polymerase iii promoters for the expression of short hairpin RNA interference molecules

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posted on 2013-01-01, 00:00 authored by B Clarke, D Cummins, K McColl, Alister WardAlister Ward, T Doran
RNA interference (RNAi) is a powerful, sequence specific, and long-lasting method of gene knockdown, and can be elicited by the expression of short-hairpin RNA (shRNA) molecules driven via polymerase III type 3 promoters from a DNA vector or transgene. To further develop RNAi as a tool in zebrafish, we have characterized the zebrafish U6 and H1 snRNA promoters and compared the efficiency of each of the promoters to express an shRNA and silence a reporter gene, relative to previously characterized U6 promoters from pufferfish, chicken, and mouse. Our results show that the zebrafish polymerase III promoters were capable of effective gene silencing in the zebrafish ZF4 cell line, but were ineffective in mammalian Vero cells. In contrast, mouse and chicken promoters were active in Vero but not ZF4 cells, highlighting the importance of homologous promoters to achieve effective silencing.

History

Journal

Zebrafish

Volume

10

Pagination

472-479

Location

New Rochelle, NY

Open access

  • Yes

ISSN

1545-8547

Language

eng

Publication classification

C1 Refereed article in a scholarly journal

Copyright notice

2013, Mary Ann Liebert

Issue

4

Publisher

Mary Ann Liebert Inc.

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