a transient transgenic rnai strategy for rapid characterization of gene function during embryonic development快速瞬态转基因rnai战略描述在胚胎发育的基因功能.pdfVIP

A Transient Transgenic RNAi Strategy for Rapid Characterization of Gene Function during Embryonic Development 1 2. 3. 1 3 Bryan C. Bjork , Yuko Fujiwara , Shannon W. Davis , Haiyan Qiu , Thomas L. Saunders , Peter 4¤ 2 3 1 Sandy , Stuart Orkin , Sally A. Camper , David R. Beier * 1 Genetics Division, Brigham Women’s Hospital, Harvard Medical School, Boston, Massachusetts, United States of America, 2 Division of Hematology and Oncology, Children’s Hospital, Harvard Medical School/Howard Hughes Medical Institute, Boston, Massachusetts, United States of America, 3 Departments of Human Genetics and Internal Medicine, University of Michigan, Ann Arbor, Michigan, United States of America, 4 The David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, United States of America 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 phenoty