デングウイルス感染蚊の網羅的遺伝子発現と新規ベクタ－コントロール

登革热病毒感染蚊子的综合基因表达和新型载体控制

• 批准号: 14J12097
• 负责人: ルントゥウェネ L.R.
• 金额: $ 1.24万
• 依托单位: The University of Tokyo (2015)Oita University (2014)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2014
• 资助国家: 日本
• 起止时间: 2014-04-25 至 2016-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-14J12097/
• 关键词: dengue virus; deep sequencing; l(2)efl; aedes aegypti; vector mosquito; RNA-seq; qRT-PCR; L(2)efl; siRNA; CCL-125; Aedes aegypti

On this fiscal year, we tried to analyze the gene in-vivo by injecting l(2)efl-1, -2, -3, and -4 dsRNAs to mosquito thoraxes and then followed by DENV-2 injection 48 hours after dsRNA-injection. We used the same dsRNA that we transfected in CCL-125. We did preliminary experiment to see how long the dsRNA would give an effect in-vivo. We found that 72 hours after dsRNA injection, the level of l(2)efl-1 to l(2)efl-5 was suppressed. Because we needed to consider that after 72 hours dsRNA might be losing its effect, we decided to inject DENV-2 48 hours after dsRNA injection. Following dsRNA-injection, mosquitoes looked healthy with only little number of mosquitoes died. Forty eight hours afterwards, we injected mosquitoes with DENV-2 and sacrificed the mosquitoes 24, 48, and 72 hours later. Unfortunately, using qRT-PCR, we found that there was no significant difference between dsRNA-injected and control mosquitoes.The reason that DENV-2 kept on replicating might because dsRNA was losing its effect at the time of DENV-2 injection. In the future, we will try to co-inject DENV-2 and dsRNA at the same time or to create a transgenic mosquito by injecting dsRNA at the embryo stage. We have submitted the results of in-vitro experiment to Japanese Journal of Infectious Disease for consideration.

在本财政年度，我们尝试通过向蚊子胸腔注射1(2)个efl-1、-2、-3和-4个dsrna，然后在注射dsrna 48小时后注射DENV-2，在体内分析该基因。我们使用了在CCL-125中转染的相同的dsRNA。我们做了初步实验，看看dsRNA在体内的作用能持续多久。我们发现，注射dsRNA 72小时后，l(2)efl-1和l(2)efl-5的水平被抑制。因为我们需要考虑到72小时后dsRNA可能会失去作用，所以我们决定在注射dsRNA 48小时后再注射DENV-2。注射dsrna后，蚊子看起来很健康，只有少量蚊子死亡。48h后分别注射DENV-2, 24h、48h、72h后处死。不幸的是，使用qRT-PCR，我们发现注射dsrna的蚊子与对照蚊子之间没有显著差异。DENV-2持续复制的原因可能是dsRNA在DENV-2注射时失去了作用。未来，我们将尝试同时注射DENV-2和dsRNA，或者在胚胎阶段通过注射dsRNA来培育转基因蚊子。我们已将体外实验结果提交日本传染病杂志审议。

项目成果

日本産Aedes (Stegomyia) scutellarisグループ蚊のチクングニアウイルス媒介能について

关于日本伊蚊（Stegomyia）盾群蚊子传播基孔肯雅病毒的能力

• 发表时间: 2015
• 作者: 福田昌子;Lucky R. Runtuwene;野口香緒里;Thipruethai Phanitchat;Raweewan Srisawat;Narumon Komalamisra;牛島廣治;倉根一郎;高崎智彦;小林隆志;江下優樹
• 通讯作者: 江下優樹

多発生硬化症の病態形成機に関わるT細胞の活性化を制御するNOD様受容体とTRAF6の役割の解明

阐明 NOD 样受体和 TRAF6 在控制多发性硬化症发病机制中 T 细胞活化中的作用

• 发表时间: 2015
• 作者: Lucky Ronald Runtuwene;Eiji Konishi;Atsushi Yamanaka;Yoshihiro Makino;Yutaka Suzuki;Tomohiko Takasaki;Ichiro Kurane;Takashi Kobayashi and Yuki Eshita;神山ながのり,飛騨野真也,林田京子,Runtuwene R. Lucky,野口かおり,園田光,江下優樹,小林隆
• 通讯作者: 神山ながのり,飛騨野真也,林田京子,Runtuwene R. Lucky,野口かおり,園田光,江下優樹,小林隆

Susceptibility of Aedes flavopictus miyarai and Aedes galloisi mosquito species in Japan to dengue type 2 virus

• DOI: 10.1016/j.apjtb.2016.03.003
• 发表时间: 2016-05-01
• 期刊: ASIAN PACIFIC JOURNAL OF TROPICAL BIOMEDICINE
• 影响因子: 1.7
• 作者: Srisawat, Raweewan;Phanitchat, Thipruethai;Eshita, Yuki
• 通讯作者: Eshita, Yuki

Potential novel anti-dengue in Aedes aegypti mosquito

埃及伊蚊的潜在新型抗登革热药物

• 发表时间: 2014
• 作者: Lucky R. Runtuwene;Shuichi Kawashima;Kaori Noguchi;Yutaka Suzuki;Sumio Sugano;Kenta Nakai;Ryuichiro Maeda;Junya Yamagishi;Chihiro Sugimoto;Tomohiko Takasaki;Ichiro Kurane;Yuki Eshita and Takashi Kobayashi
• 通讯作者: Yuki Eshita and Takashi Kobayashi

日本国内への侵入が危惧される節足動物媒介性ウイルスと日本の蚊について

关于担心入侵日本的节肢动物传播病毒和日本蚊子

• 发表时间: 2015
• 作者: 江下優樹,福田昌子,Lucky R. Runtuwene,野口香緒里,飛弾野真也,神山長慶,林田京子,小林隆志;Thipruethai Phanitchat,Raweewan Srisawat,Narumon Komalamisra,成田弘成,牛島廣治,倉根一郎,高崎智彦
• 通讯作者: Thipruethai Phanitchat,Raweewan Srisawat,Narumon Komalamisra,成田弘成,牛島廣治,倉根一郎,高崎智彦