[AalDV-3介导的mirRNA海绵传递系统的构建及其对埃及伊蚊细胞和幼虫的影响]。

Jinghua Guo, Yanhai Wang
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引用次数: 0

摘要

microRNAs (miRNAs)不仅在蚊子的生长发育调控中起着关键作用,而且在病原与媒介的相互作用中也起着重要作用。因此,mirna可以作为分子靶点,开发一种蚊虫和蚊媒疾病控制的替代方法。然而,仍然需要一个有效的交付系统。蚊子致密病毒作为表达外源毒素或小干扰rna分子的转导剂,在体内和体外均具有控制媒介的潜力。在本研究中,我们利用内含子miRNA海绵表达策略,建立了重组白纹伊蚊致密病毒-3(AalDV-3) miRNA海绵表达系统。为了检测重组病毒给药miRNA海绵对内源性miR-210的抑制作用,分别感染埃及伊蚊Aag2细胞和1 -2幼虫。利用内含子跨引物RT-PCR检测内含子miRNA海绵表达构建体在体外和体内的剪接情况,并通过qPCR确认miR-210的相对表达水平。结果表明,aaldv -3在体外和体内均可通过产生反义海绵来减少内源性miRNAs,为蚊虫基因功能分析提供了工具,为致密病毒在媒介控制中的应用奠定了基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Construction of AalDV-3 Mediated mirRNA Sponge Delivery System and Its Effect on Cell and Larva of Aedes aegypti].

microRNAs (miRNAs) not only play the key roles in regulation of the growth and development of mosquito, but also has an important function in interaction between the pathogen and vector. So, miRNAs can be used as the molecular target for development of an alternative method for mosquito and mosquito-borne disease control. However, an effective delivery system is still need. Mosquito densovirus have the potential for vector control as transducing agents to express foreign toxins or small interfering RNAs molecules in vitro and in vivo. In this study, we report the development of a recombinant Aedes albopictus densovirus-3(AalDV-3) miRNA sponge expression system, using an intronic miRNA sponge expression strategy. To test the inhibition effect of recombinant virus medicated miRNA sponge on endogenous miR-210, Aedes aegypti Aag2 cell and 1st-2nd larvae were infected, respectively. The splicing of the intronic miRNA sponge expression constructs in vitro and in vivo were tested by RT-PCR with intron span primers, and the relatively expression level of miR-210 was confirmed by qPCR. As results,AalDV-3can be used to decrease endogenous miRNAs by generating an antisense sponge in vitro and in vivo, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of densovirus for vector control.

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