| A rapid and scalable system for studying gene function in mice using conditional RNA interference. | |
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MedLine Citation:
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PMID: 21458673 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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RNA interference is a powerful tool for studying gene function, however, the reproducible generation of RNAi transgenic mice remains a significant limitation. By combining optimized fluorescence-coupled miR30-based shRNAs with high efficiency ES cell targeting, we developed a fast, scalable pipeline for the production of shRNA transgenic mice. Using this system, we generated eight tet-regulated shRNA transgenic lines targeting Firefly and Renilla luciferases, Oct4 and tumor suppressors p53, p16(INK4a), p19(ARF) and APC and demonstrate potent gene silencing and GFP-tracked knockdown in a broad range of tissues in vivo. Further, using an shRNA targeting APC, we illustrate how this approach can identify predicted phenotypes and also unknown functions for a well-studied gene. In addition, through regulated gene silencing we validate APC/Wnt and p19(ARF) as potential therapeutic targets in T cell acute lymphoblastic leukemia/lymphoma and lung adenocarcinoma, respectively. This system provides a cost-effective and scalable platform for the production of RNAi transgenic mice targeting any mammalian gene. PAPERCLIP: |
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Authors:
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Prem K Premsrirut; Lukas E Dow; Sang Yong Kim; Matthew Camiolo; Colin D Malone; Cornelius Miething; Claudio Scuoppo; Johannes Zuber; Ross A Dickins; Scott C Kogan; Kenneth R Shroyer; Raffaella Sordella; Gregory J Hannon; Scott W Lowe |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Cell Volume: 145 ISSN: 1097-4172 ISO Abbreviation: Cell Publication Date: 2011 Apr |
Date Detail:
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Created Date: 2011-04-04 Completed Date: 2011-05-27 Revised Date: 2012-09-19 |
Medline Journal Info:
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Nlm Unique ID: 0413066 Medline TA: Cell Country: United States |
Other Details:
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Languages: eng Pagination: 145-58 Citation Subset: IM |
Copyright Information:
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Copyright © 2011 Elsevier Inc. All rights reserved. |
Affiliation:
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Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Adenocarcinoma
/
genetics,
therapy Animals Embryonic Stem Cells / metabolism Gene Knockdown Techniques / economics, methods* Lung Neoplasms / genetics, therapy Mice Mice, Transgenic MicroRNAs / genetics Precursor T-Cell Lymphoblastic Leukemia-Lymphoma / genetics, therapy RNA Interference* RNA Processing, Post-Transcriptional RNA, Small Interfering / genetics Signal Transduction Wnt Proteins / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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P01 CA013106-40/CA/NCI NIH HHS; R01 GM062534-12/GM/NIGMS NIH HHS; //Howard Hughes Medical Institute; //Howard Hughes Medical Institute |
| Chemical | |
Reg. No./Substance:
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0/MicroRNAs; 0/RNA, Small Interfering; 0/Wnt Proteins |
| Comments/Corrections | |
Comment In:
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Nat Methods. 2011 May;8(5):374
[PMID:
21678621
]
Nat Rev Genet. 2011 Jun;12(6):380 [PMID: 21577222 ] |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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