Clemons Anthony, Haugen Morgan, Severson David, Duman-Scheel Molly
Department of Medical and Molecular Genetics, Indiana University School of Medicine, South Bend, IN 46617, USA.
Cold Spring Harb Protoc. 2010 Oct 1;2010(10):pdb.prot5511. doi: 10.1101/pdb.prot5511.
Blood-feeding mosquitoes, including the dengue and yellow fever vector Aedes aegypti, transmit many of the world's deadliest diseases. Such diseases have resurged in developing countries and pose clear threats for epidemic outbreaks in developed countries. Recent mosquito genome projects have stimulated interest in the potential for arthropod-borne disease control by genetic manipulation of vector insects, and genes that regulate development are of particular interest. In recent years, RNA interference (RNAi) has proven to be an effective strategy for inhibiting gene function in many organisms. This protocol describes a method for knockdown of embryonic genes in Ae. aegypti embryos by microinjection of small interfering RNA (siRNA) designed to target a specific gene of interest. The procedure includes a strategy for siRNA design, microinjection, and measurement of knockdown effectiveness.
吸食血液的蚊子，包括登革热和黄热病的传播媒介埃及伊蚊，传播着世界上许多最致命的疾病。这些疾病在发展中国家再度出现，并对发达国家的疫情爆发构成了明显威胁。最近的蚊子基因组项目激发了人们对通过基因操纵媒介昆虫来控制节肢动物传播疾病的潜力的兴趣，而调控发育的基因尤其受到关注。近年来，RNA干扰（RNAi）已被证明是在许多生物体中抑制基因功能的有效策略。本方案描述了一种通过显微注射针对特定目标基因设计的小干扰RNA（siRNA）来敲低埃及伊蚊胚胎中胚胎基因的方法。该程序包括siRNA设计、显微注射和敲低有效性测量的策略。
