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A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development

Author(s)
Bjork, Bryan C.; Fujiwara, Yuko; Davis, Shannon W.; Qiu, Haiyan; Saunders, Thomas L.; Sandy, Peter; Orkin, Stuart H.; Camper, Sally A.; Beier, David R.; ... Show more Show less
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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.
Date issued
2010-12
URI
http://hdl.handle.net/1721.1/63160
Department
Koch Institute for Integrative Cancer Research at MIT
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
Bjork BC, Fujiwara Y, Davis SW, Qiu H, Saunders TL, et al. (2010) A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development. PLoS ONE 5(12): e14375. doi:10.1371/journal.pone.0014375
Version: Final published version
ISSN
1932-6203

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