Transgene-induced cell death following dengue-2 virus infection in Aedes aegypti.
Transgene-induced cell death following dengue-2 virus infection in Aedes aegypti.
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DOI：
10.1038/s41598-023-32895-9
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发表时间：
2023-04-12
影响因子：
4.6
通讯作者：
Capurro ML
中科院分区：
文献类型：
--
作者：
Carvalho DO;Costa-da-Silva AL;Petersen V;de Souza MS;Ioshino RS;Marques ICS;Franz AWE;Olson KE;James AA;Capurro ML
登革热病毒（DENV）是蚊媒黄病毒，每年造成数百万人感染，对全球公共卫生系统构成挑战。埃及伊蚊是向人类传播DENV的主要媒介物种。控制Ae。由于繁殖地丰富和病媒种群对杀虫剂的抗药性日益增加，埃及伊蚊的传播比较困难。制定新的病媒控制策略对于减少疾病负担至关重要。一种潜在的方法是在基因上取代Ae。埃及人群体与对DENV传播具有高度抗性的媒介群体。在这里，我们专注于在基因修饰的Ae中产生登革2型病毒（DENV-2）抗性的替代策略。埃及伊蚊，其中蚊子表达非活性形式的Michelob_x（Mx），一种细胞凋亡抑制剂（IAP）的拮抗剂，以诱导存在活跃复制的DENV-2的那些细胞中的细胞凋亡。Mx的失活形式的侧翼是RRRRSAG切割基序，其被感染的DENV-2的NS 2B/NS 3蛋白酶识别，从而释放和激活Mx，然后诱导细胞凋亡。当感染DENV-2时，我们的转基因株表现出比非转基因对照显著更高的死亡率。我们还将含有与eGFP融合的失活Mx的DNA构建体转染到C6/36蚊子细胞中，并在DENV-2感染后第3天和第6天间接观察到Mx活化。有明显的迹象表明病毒NS 2B/NS 3蛋白酶切割转基因，从而将Mx蛋白释放到细胞质中，如通过检测感染细胞中的eGFP表达所证实的。本研究证明了病毒感染可用于诱导受感染蚊子细胞凋亡的概念。
Dengue viruses (DENVs) are mosquito-borne flaviviruses causing millions of human infections each year and pose a challenge for public health systems worldwide. Aedes aegypti is the principal vector species transmitting DENVs to humans. Controlling Ae. aegypti is difficult due to the abundance of breeding sites and increasing insecticide resistance in the vector populations. Developing new vector control strategies is critical for decreasing the disease burden. One potential approach is genetically replacing Ae. aegypti populations with vector populations highly resistant to DENV transmission. Here, we focus on an alternative strategy for generating dengue 2 virus (DENV-2) resistance in genetically-modified Ae. aegypti in which the mosquitoes express an inactive form of Michelob_x (Mx), an antagonist of the Inhibitor of Apoptosis (IAP), to induce apoptosis in those cells in which actively replicating DENV-2 is present. The inactive form of Mx was flanked by the RRRRSAG cleavage motif, which was recognized by the NS2B/NS3 protease of the infecting DENV-2 thereby releasing and activating Mx which then induced apoptosis. Our transgenic strain exhibited a significantly higher mortality rate than the non-transgenic control when infected with DENV-2. We also transfected a DNA construct containing inactive Mx fused to eGFP into C6/36 mosquito cells and indirectly observed Mx activation on days 3 and 6 post-DENV-2 infections. There were clear signs that the viral NS2B/NS3 protease cleaved the transgene, thereby releasing Mx protein into the cytoplasm, as was confirmed by the detection of eGFP expression in infected cells. The present study represents proof of the concept that virus infection can be used to induce apoptosis in infected mosquito cells.
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DOI：
10.1098/rspb.2008.0846
发表时间：
2008-12-22
影响因子：
4.7
作者：
Gould, Fred;Huang, Yunxin;Lloyd, Alun L.
通讯作者：
Lloyd, Alun L.
影响因子：
64.8
作者：
Gogala, Marko;Becker, Thomas;Beckmann, Roland
通讯作者：
Beckmann, Roland
影响因子：
3.8
作者：
Franz AW;Sanchez-Vargas I;Raban RR;Black WC 4th;James AA;Olson KE
通讯作者：
Olson KE
影响因子：
3.8
作者：
Coates, CJ;Jasinskiene, N;James, AA
通讯作者：
James, AA
影响因子：
6.7
作者：
Buchman, Anna;Gamez, Stephanie;Akbari, Omar S.
通讯作者：
Akbari, Omar S.