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  An inducible RNA interference system for the functional dissection of mouse embryogenesis.

Vidigal, J. A., Morkel, M., Wittler, L., Brouwer-Lehmitz, A., Grote, P., Macura, K., et al. (2010). An inducible RNA interference system for the functional dissection of mouse embryogenesis. Nucleic Acids Research, 38(11), e122-e122. doi:10.1093/nar/gkq199.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7B81-3 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-7B84-E
Genre: Journal Article
Alternative Title : Nucleic Acids Res.

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 Creators:
Vidigal, Joana A.1, Author
Morkel, Markus2, Author              
Wittler, Lars3, Author              
Brouwer-Lehmitz, Antje2, Author              
Grote, Phillip2, Author              
Macura, Karol2, Author              
Herrmann, Bernhard G.2, Author              
Affiliations:
1Max Planck Society, ou_persistent13              
2Dept. of Developmental Genetics (Head: Bernhard G. Herrmann), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433548              
3Transgene Unit (Head: Lars Wittler), Scientific Service (Head: Manuela B. Urban), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1479663              

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 Abstract: Functional analysis of multiple genes is key to understanding gene regulatory networks controlling embryonic development. We have developed an integrated vector system for inducible gene silencing by shRNAmir-mediated RNA interference in mouse embryos, as a fast method for dissecting mammalian gene function. For validation of the vector system, we generated mutant phenotypes for Brachyury, Foxa2 and Noto, transcription factors which play pivotal roles in embryonic development. Using a series of Brachyury shRNAmir vectors of various strengths we generated hypomorphic and loss of function phenotypes allowing the identification of Brachyury target genes involved in trunk development. We also demonstrate temporal control of gene silencing, thus bypassing early embryonic lethality. Importantly, off-target effects of shRNAmir expression were not detectable. Taken together, the system allows the dissection of gene function at unprecedented detail and speed, and provides tight control of the genetic background minimizing intrinsic variation.

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Language(s): eng - English
 Dates: 2010-03-27
 Publication Status: Published in print
 Pages: -
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 Identifiers: eDoc: 536196
DOI: 10.1093/nar/gkq199
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Title: Nucleic Acids Research
  Alternative Title : Nucleic Acids Res.
Source Genre: Journal
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Pages: - Volume / Issue: 38 (11) Sequence Number: - Start / End Page: e122 - e122 Identifier: ISSN: 0305-1048