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- Title
Gene silencing in Xenopus laevis by DNA vector-based RNA interference and transgenesis.
- Authors
Ming Li; Rohrer, Baerbel
- Abstract
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.Cell Research (2006) 16: 99–105. doi:10.1038/sj.cr.7310013; published online 16 January 2006
- Publication
Cell Research, 2006, Vol 16, Issue 1, p99
- ISSN
1001-0602
- Publication type
Academic Journal
- DOI
10.1038/sj.cr.7310013