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

Clarke, Brian D, Cummins, David M, McColl, Ken A, Ward, Alister C and Doran, Tim J 2013, Characterization of zebrafish polymerase iii promoters for the expression of short hairpin RNA interference molecules, Zebrafish, vol. 10, no. 4, pp. 472-479.

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Title Characterization of zebrafish polymerase iii promoters for the expression of short hairpin RNA interference molecules
Author(s) Clarke, Brian D
Cummins, David M
McColl, Ken A
Ward, Alister C
Doran, Tim J
Journal name Zebrafish
Volume number 10
Issue number 4
Start page 472
End page 479
Total pages 8
Publisher Mary Ann Liebert Inc.
Place of publication New Rochelle, NY
Publication date 2013
ISSN 1545-8547
Summary 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.
Language eng
Field of Research 060403 Developmental Genetics (incl Sex Determination)
Socio Economic Objective 970106 Expanding Knowledge in the Biological Sciences
HERDC Research category C1 Refereed article in a scholarly journal
Copyright notice ©2013, Mary Ann Liebert
Persistent URL http://hdl.handle.net/10536/DRO/DU:30059499

Document type: Journal Article
Collections: School of Medicine
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Every reasonable effort has been made to ensure that permission has been obtained for items included in DRO. If you believe that your rights have been infringed by this repository, please contact drosupport@deakin.edu.au.