Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/62061
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Type: Journal article
Title: Robust, reversible gene knockdown using a single lentiviral short hairpin RNA vector
Author: Brown, C.
Sadlon, T.
Gargett, T.
Melville, E.
Zhang, R.
Drabsch, Y.
Ling, M.
Strathdee, C.
Gonda, T.
Barry, S.
Citation: Human Gene Therapy, 2010; 21(8):1005-1017
Publisher: Mary Ann Liebert Inc
Issue Date: 2010
ISSN: 1043-0342
1557-7422
Statement of
Responsibility: 
Cheryl Y. Brown, Timothy Sadlon, Tessa Gargett, Elizabeth Melville, Rui Zhang, Yvette Drabsch, Michael Ling, Craig A. Strathdee, Thomas J. Gonda, and Simon C. Barry
Abstract: Manipulation of gene expression is an invaluable tool to study gene function in vitro and in vivo. The application of small inhibitory RNAs to knock down gene expression provides a relatively simple, elegant, but transient approach to study gene function in many cell types as well as in whole animals. Short hairpin structures (shRNAs) are a logical advance as they can be expressed continuously and are hence suitable for stable gene knockdown. Drug-inducible systems have now been developed; however, application of the technology has been hampered by persistent problems with low or transient expression, leakiness or poor inducibility of the short hairpin, and lack of reversibility. We have developed a robust, versatile, single lentiviral vector tool that delivers tightly regulated, fully reversible, doxycycline-responsive knockdown of target genes (FOXP3 and MYB), using single short hairpin RNAs. To demonstrate the capabilities of the vector we targeted FOXP3 because it plays a critical role in the development and function of regulatory T cells. We also targeted MYB because of its essential role in hematopoiesis and implication in breast cancer progression. The versatility of this vector is hence demonstrated by knockdown of distinct genes in two biologically separate systems.
Keywords: Animals; Humans; Mice; Lentivirus; Doxycycline; Proto-Oncogene Proteins c-myb; RNA, Small Interfering; Gene Targeting; Transfection; Gene Expression; Genetic Vectors; Forkhead Transcription Factors; Gene Knockdown Techniques; HEK293 Cells
Rights: Copyright Mary Ann Liebert, Inc.
RMID: 0020100686
DOI: 10.1089/hum.2009.107
Appears in Collections:Paediatrics publications

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