Robust, reversible gene knockdown using a single lentiviral short hairpin RNA vector

Brown, Cheryl Y., Sadlon, Timothy, Gargett, Tessa, Melville, Elizabeth, Zhang, Rui, Drabsch, Yvette, Ling, Michael, Strathdee, Craig A., Gonda, Thomas J. and Barry, Simon C. (2010) Robust, reversible gene knockdown using a single lentiviral short hairpin RNA vector. Human Gene Therapy, 21 8: 1005-1017. doi:10.1089/hum.2009.107

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Author Brown, Cheryl Y.
Sadlon, Timothy
Gargett, Tessa
Melville, Elizabeth
Zhang, Rui
Drabsch, Yvette
Ling, Michael
Strathdee, Craig A.
Gonda, Thomas J.
Barry, Simon C.
Title Robust, reversible gene knockdown using a single lentiviral short hairpin RNA vector
Journal name Human Gene Therapy   Check publisher's open access policy
ISSN 1043-0342
1557-7422
Publication date 2010-08-01
Sub-type Article (original research)
DOI 10.1089/hum.2009.107
Open Access Status File (Author Post-print)
Volume 21
Issue 8
Start page 1005
End page 1017
Total pages 13
Place of publication New Rochelle, NY, United States
Publisher Mary Ann Liebert
Collection year 2011
Language eng
Formatted 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.
Keyword Regulatory T-cells
G-CSF delivery
C-MYB
Transgene expression
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2011 Collection
UQ Diamantina Institute - Open Access Collection
UQ Diamantina Institute Publications
 
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Citation counts: TR Web of Science Citation Count  Cited 14 times in Thomson Reuters Web of Science Article | Citations
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Created: Sun, 29 Aug 2010, 10:06:06 EST