Splice-activated UAS hairpin vector gives complete RNAi knockout of single or double target transcripts in Drosophila melanogaster
Splice-activated UAS hairpin vector gives complete RNAi knockout of single or double target transcripts in Drosophila melanogaster
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DOI：
10.1002/gene.10122
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发表时间：
2002-09-01
期刊：
影响因子：
1.5
通讯作者：
Gubb, D
中科院分区：
文献类型：
--
作者：
Reichhart, JM;Ligoxygakis, P;Gubb, D
描述了一种提供nec的完全RNAi敲除的DNA载体。稳定的发夹环是无活性的，但内含子间隔区的引入激活了RNAi反应。叉状、Drs和BG4转录物的类似构建体也提供靶向抑制。此外，“剪接激活”可影响双敲除载体中的两个触发序列。我们认为，剪接机制可能会导致局部的“熔化”的RNA，然后再退火。这个过程将消除在生理条件下互补RNA链杂交所预期的错配区域。与植物不同的是，我们发现，
A DNA vector giving complete RNAi knockout of nec is described. A stable hairpin-loop is inactive, but introduction of an intronic spacer activates the RNAi response. Similar constructs for the forked, Drs, and BG4 transcripts also give targeted suppression. Furthermore,“splice activation” can affect two trigger sequences, in a double knockout vector. We suggest that the splicing mechanism might cause localized “melting” of RNA followed by reannealing. This process would eliminate mismatched regions expected from hybridization of complementary RNA strands under physiological conditions. Unlike in plants, we show that the RNAi response in