A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development
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A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development
| Title: | A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development |
| Author: |
Davis, Shannon W.; Qiu, Haiyan; Saunders, Thomas L.; Sandy, Peter; Camper, Sally A.; Bjork, Bryan Cary; Fujiwara, Yuko; Orkin, Stuart Holland; Beier, David Randolph
Note: Order does not necessarily reflect citation order of authors. |
| Citation: | Bjork, Bryan C., Yuko Fujiwara, Shannon W. Davis, Haiyan Qiu, Thomas L. Saunders, Peter Sandy, Stuart Orkin, Sally A. Camper, and David R. Beier. 2010. A transient transgenic RNAi strategy for rapid characterization of gene function during embryonic development. PLoS ONE 5(12): e14375. |
| Full Text & Related Files: |
3002952.pdf (2.287Mb; PDF) 
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| Abstract: | RNA interference (RNAi) is a powerful strategy for studying the phenotypic consequences of reduced gene expression levels in model systems. To develop a method for the rapid characterization of the developmental consequences of gene dysregulation, we tested the use of RNAi for “transient transgenic” knockdown of mRNA in mouse embryos. These methods included lentiviral infection as well as transposition using the Sleeping Beauty (SB) and PiggyBac (PB) transposable element systems. This approach can be useful for phenotypic validation of putative mutant loci, as we demonstrate by confirming that knockdown of Prdm16 phenocopies the ENU-induced cleft palate (CP) mutant, csp1. This strategy is attractive as an alternative to gene targeting in embryonic stem cells, as it is simple and yields phenotypic information in a matter of weeks. Of the three methodologies tested, the PB transposon system produced high numbers of transgenic embryos with the expected phenotype, demonstrating its utility as a screening method. |
| Published Version: | doi:10.1371/journal.pone.0014375 |
| Other Sources: | http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3002952/pdf/ |
| Terms of Use: | This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAA |
| Citable link to this page: | http://nrs.harvard.edu/urn-3:HUL.InstRepos:4874766 |
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