Transgene-mediated suppression of the RNA interference pathway in Aedes aegypti interferes with gene silencing and enhances Sindbis virus and dengue virus type 2 replication.
Transgene-mediated suppression of the RNA interference pathway in Aedes aegypti interferes with gene silencing and enhances Sindbis virus and dengue virus type 2 replication.
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DOI：
10.1111/imb.12008
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发表时间：
2013-02
影响因子：
2.6
通讯作者：
Franz AW
中科院分区：
文献类型：
--
作者：
Khoo CC;Doty JB;Heersink MS;Olson KE;Franz AW
RNA干扰（RNAi）是埃及伊蚊体内主要的先天性抗病毒途径，可应答复制的虫媒病毒，如DENV和SINV。蚊子的RNAi机制能够完全消除来自Ae的DENV 2。埃及人。另一方面，RNAi途径的关键基因的瞬时沉默增加了SINV和DENV 2的复制，允许病毒以更高的速率暂时克服剂量依赖性中肠感染和逃逸屏障。在这里，我们表达FHV-B2从多聚泛素（PUb）启动子在Ae。使用ΦC31定点重组系统研究转基因介导的RNAi途径抑制对SINV-TR 339 eGFP和DENV 2-QR 94感染的影响，后者已显示在埃及伊蚊中面临强中肠逃逸屏障（MEB）。埃及人。FHV-B2在转基因系PUbB 2 P61的糖和血喂养的蚊子的中肠中组成型表达。B2过表达抑制PUbB 2 P61蚊子中肠组织中羧肽酶A-1（AeCPA-1）的RNA沉默。在用SINV-TR 339 eGFP或DENV 2-QR 94口服攻击后，在血餐后7天（pbm），PUbB 2 P61雌性的中肠中的平均滴度显著高于非转基因对照蚊子中的平均滴度。在14天pbm时，在用SINV-TR 339 eGFP感染的PubB 2 P61蚊子中尸体的感染率显著增加。在用DENV 2-QR 94感染后，在14天pbm时，表达B2的蚊子中中肠感染率显著增加。然而，PUbB 2 P61中的B2表达不增加DENV 2-QR 94传播率，表明感染表型主要不由RNAi控制。
RNA interference (RNAi) is the major innate antiviral pathway in Aedes aegypti that responds to replicating arboviruses such as DENV and SINV. The mosquito’s RNAi machinery is capable of completely eliminating DENV2 from Ae. aegypti. On the other hand, transient silencing of key genes of the RNAi pathway increases replication of SINV and DENV2, allowing the viruses to temporally overcome dose-dependent midgut infection and –escape barriers at higher rates. Here we expressed FHV-B2 from the poly-ubiquitin (PUb) promoter in Ae. aegypti using the ΦC31 site-directed recombination system to investigate the impact of transgene-mediated RNAi pathway suppression on infections with SINV-TR339eGFP and DENV2-QR94, the latter of which has been shown to be confronted with a strong midgut escape barrier (MEB) in Ae. aegypti. FHV-B2 was constitutively expressed in midguts of sugar- and bloodfed mosquitoes of transgenic line PUbB2 P61. B2 over-expression suppressed RNA silencing of carboxypeptidase A-1 (AeCPA-1) in midgut tissue of PUbB2 P61 mosquitoes. Following oral challenge with SINV-TR339eGFP or DENV2-QR94, mean titers in midguts of PUbB2 P61 females were significantly higher at 7 days post-bloodmeal (pbm) than in those of non-transgenic control mosquitoes. At 14 days pbm, infection rates of carcasses were significantly increased in PubB2 P61 mosquitoes infected with SINV-TR339eGFP. Following infection with DENV2-QR94, midgut infection rates were significantly increased in the B2-expressing mosquitoes at 14 days pbm. However, B2 expression in PUbB2 P61 did not increase the DENV2-QR94 dissemination rate, indicating that the infection phenotype was not primarily controlled by RNAi.
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影响因子：
4.2
作者：
Campbell CL;Keene KM;Brackney DE;Olson KE;Blair CD;Wilusz J;Foy BD
通讯作者：
Foy BD
影响因子：
2.6
作者：
Mathur, G.;Sanchez-Vargas, I.;James, A. A.
通讯作者：
James, A. A.
影响因子：
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作者：
Sanchez-Vargas, I;Travanty, EA;Olson, KE
通讯作者：
Olson, KE
影响因子：
4.2
作者：
Khoo CC;Piper J;Sanchez-Vargas I;Olson KE;Franz AW
通讯作者：
Franz AW
影响因子：
6.7
作者：
Sanchez-Vargas, Irma;Scott, Jaclyn C.;Poole-Smith, B. Katherine;Franz, Alexander W. E.;Barbosa-Solomieu, Valerie;Wilusz, Jeffrey;Olson, Ken E.;Blair, Carol D.
通讯作者：
Blair, Carol D.