A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development
作者： Bryan C. Bjork , Yuko Fujiwara , Shannon W. Davis , Haiyan Qiu , Thomas L. Saunders
DOI: 10.1371/JOURNAL.PONE.0014375
关键词: Transgene 、 Gene expression 、 Genetics 、 RNA interference 、 Gene 、 Biology 、 Gene targeting 、 Transposable element 、 Gene knockdown 、 Mutant 、 General Biochemistry, Genetics and Molecular Biology 、 General Agricultural and Biological Sciences 、 General Medicine
摘要: RNA interference (RNAi) is a powerful strategy for studying the phenotypic consequences of reduced gene expression levels in model systems. To develop method rapid characterization developmental dysregulation, we tested use RNAi ‘‘transient transgenic’’ knockdown mRNA mouse embryos. These methods included lentiviral infection as well transposition using Sleeping Beauty (SB) and PiggyBac (PB) transposable element This approach can be useful validation putative mutant loci, demonstrate by confirming that Prdm16 phenocopies ENU-induced cleft palate (CP) mutant, csp1. attractive an alternative to targeting embryonic stem cells, it simple yields information matter weeks. Of three methodologies tested, PB transposon system produced high numbers transgenic embryos with expected phenotype, demonstrating its utility screening method.
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