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首页> 外文期刊>Cell Research >Gene silencing in Xenopus laevis by DNA vector-based RNA interference and transgenesis.
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Gene silencing in Xenopus laevis by DNA vector-based RNA interference and transgenesis.

机译：通过基于DNA载体的RNA干扰和转基因，非洲爪蟾中的基因沉默。

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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 approximately 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.
机译：使用非洲爪蟾U6启动子开发了基于载体的RNAi表达系统，该启动子通过RNA聚合酶III转录小RNA基因。该系统首先在非洲爪蟾（Xenopus laevis）细胞系中得到验证，设计了对GFP基因具有特异性的短发夹DNA。将基于载体的RNAi和GFP基因共转染到非洲爪蟾XR1细胞中，显着减少了表达GFP的细胞数量和整体GFP荧光。随后在GFP转基因非洲爪蟾胚胎中验证了基于载体的RNAi。来自GFP转基因雄性的精子核和RNAi构建体孵育的精子核分别用于受精。与对照相比，这些转基因胚胎中的RNAi使GFP mRNA和蛋白质减少了约60％。这种转基因驱动的RNAi在抑制非洲爪蟾转基因株系中的GFP表达方面具有特异性和稳定性。通过基于载体的RNAi和非洲爪蟾转基因进行的基因沉默可能为脊椎动物模型系统中的``基因功能抑制''研究提供替代方法.Cell Research（2006）16：99-105。 doi：10.1038 / sj.cr.7310013;在线发布于2006年1月16日。

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