Chitosan/interfering RNA nanoparticle mediated gene silencing in disease vector mosquito larvae.
Chitosan/interfering RNA nanoparticle mediated gene silencing in disease vector mosquito larvae.
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DOI：
10.3791/52523
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发表时间：
2015-03-25
期刊：
影响因子：
--
通讯作者：
Duman-Scheel M
中科院分区：
文献类型：
--
作者：
Zhang X;Mysore K;Flannery E;Michel K;Severson DW;Zhu KY;Duman-Scheel M
在这里，我们描述了一个程序，通过使用壳聚糖/干扰RNA纳米颗粒，幼虫摄入的疾病载体蚊子抑制基因功能。病媒蚊子给人类带来的痛苦比其他任何生物都多，每年造成100多万人死亡。蚊子基因组项目促进了对蚊子生物学新方面的研究，包括对非洲主要疟疾媒介冈比亚按蚊和登革热和黄热病媒介埃及伊蚊的功能遗传研究。RNA干扰（RNAi）介导的基因沉默已被用于靶向这两种疾病载体蚊子物种中的感兴趣的基因。在这里，我们描述了一个程序，用于制备壳聚糖/干扰RNA纳米颗粒，结合食物和幼虫摄入。这种技术上简单、高通量和相对便宜的方法，与长双链RNA（dsRNA）或小干扰RNA（siRNA）分子相容，已用于成功敲除A. gambiae和A.埃及伊蚊幼虫在幼虫喂食后，通过qRT-PCR或原位杂交验证的敲除可以至少持续到蛹后期。这一方法可适用于各种各样的蚊子和其他昆虫物种，包括农业害虫，以及其他非模式生物。除了在研究实验室中的实用性外，在未来，壳聚糖，一种廉价，无毒和可生物降解的聚合物，可能会被用于该领域。
Here we describe a procedure for inhibiting gene function in disease vector mosquitoes through the use of chitosan/interfering RNA nanoparticles that are ingested by larvae. Vector mosquitoes inflict more human suffering than any other organism—and kill more than one million people each year. The mosquito genome projects facilitated research in new facets of mosquito biology, including functional genetic studies in the primary African malaria vector Anopheles gambiae and the dengue and yellow fever vector Aedes aegypti. RNA interference- (RNAi-) mediated gene silencing has been used to target genes of interest in both of these disease vector mosquito species. Here, we describe a procedure for preparation of chitosan/interfering RNA nanoparticles that are combined with food and ingested by larvae. This technically straightforward, high-throughput, and relatively inexpensive methodology, which is compatible with long double stranded RNA (dsRNA) or small interfering RNA (siRNA) molecules, has been used for the successful knockdown of a number of different genes in A. gambiae and A. aegypti larvae. Following larval feedings, knockdown, which is verified through qRT-PCR or in situ hybridization, can persist at least through the late pupal stage. This methodology may be applicable to a wide variety of mosquito and other insect species, including agricultural pests, as well as other non-model organisms. In addition to its utility in the research laboratory, in the future, chitosan, an inexpensive, non-toxic and biodegradable polymer, could potentially be utilized in the field.
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影响因子：
--
作者：
Mysore K;Andrews E;Li P;Duman-Scheel M
通讯作者：
Duman-Scheel M
DOI：
10.1073/pnas.1000251107
发表时间：
2010-03-09
影响因子：
11.1
作者：
Fu, Guoliang;Lees, Rosemary S.;Alphey, Luke
通讯作者：
Alphey, Luke
影响因子：
17.3
作者：
Gaj, Thomas;Gersbach, Charles A.;Barbas, Carlos F., III
通讯作者：
Barbas, Carlos F., III
荧光纳米颗粒传递双链RNA以实现害虫的遗传控制
影响因子：
29.4
作者：
He, Bicheng;Chu, Yuan;Shen, Jie
通讯作者：
Shen, Jie
影响因子：
2.1
作者：
Lopez-Martinez, Giancarlo;Meuti, Megan;Denlinger, David L.
通讯作者：
Denlinger, David L.