Please use this identifier to cite or link to this item:
https://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. |
DOI: | 10.1089/hum.2009.107 |
Published version: | http://dx.doi.org/10.1089/hum.2009.107 |
Appears in Collections: | Aurora harvest Paediatrics publications |
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hdl_62061.pdf | Accepted version | 930.77 kB | Adobe PDF | View/Open |
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