Gene silencing in Xenopus laevis by DNA vector-based RNA interference and transgenesis  

作　　者：Ming Li Baerbel Rohrer 

机构地区：[1]Departments of Neurosciences , Division of Research, Medical University of South Carolina, 167 Ashley Ave, SEI 511, Charleston, SC 29425, USA [2]Departments of Ophthalmology, Division of Research, Medical University of South Carolina, 167 Ashley Ave, SEI 511, Charleston, SC 29425, USA

出　　处：《Cell Research》2006年第1期99-105,共7页细胞研究（英文版）

摘　　要：A vector-based RNAi expression system was developed using the Xenopus tropicalis U6 promoter, which transcribes small RNA genes by RNA polymerase Ⅲ. The system was first validated in a Xenopus laevis cell line, designing a short hairpin DNA specific for the GFP gene. Co-transfection of the vector-based RNAi and the GFP gene into Xenopus XR1 cells significantly decreased the number of GFP-expressing cells and overall GFP fluorescence. Vector-based RNAi was subsequently validated in GFP transgenic Xenopus embryos. Sperm nuclei from GFP transgenic males and RNAi construct-incubated-sperm nuclei were used for fertilization, respectively. GFP mRNA and protein were reduced by -60% by RNAi in these transgenic embryos compared with the control. This transgene-driven RNAi is specific and stable in inhibiting GFP expression in the Xenopus laevis transgenic line. Gene silencing by vector-based RNAi and Xenopus transgenesis may provide an alternative for ＇repression of gene function＇ studies in vertebrate model systems.

关 键 词：RNAi Xenopus U6 promoter TRANSGENIC Xenopus laevis GFP 

分 类 号：R394[医药卫生—医学遗传学]