Elucidation of mosquito-malaria parasite interaction using genetically engineered mosquitoes

使用基因工程蚊子阐明蚊子与疟疾寄生虫的相互作用

• 批准号: 16590345
• 负责人: YOSHIDA Shigeto
• 金额: $ 2.11万
• 依托单位: JICHI MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590345/
• 关键词: malaria; anopheline mosquito; transgenic; salivary gland; CEL-III; Plasmodium; Anopheles; lectin; スポロゾイト

Malaria sporozoites invade the mosquito salivary glands and wait in the salivary duct until the next blood feeding. The mechanisms of the process and molecules involved in the salivary gland invasion remain largely unknown. To establish a robust salivary gland-specific transgene expression in Anopheles stephensi, we obtained a salivary gland-specific promoter for a gene encoding Anopheles anti-platelet aggregation protein (AAPP). The aapp promoter is a female salivary gland-specific and blood meal-inducible strong promoter. Using this promoter, we generated a transgenic An.stephensi expressing abundant Discosoma sp. red fluorescent protein (DsRed) in the distal-lateral lobes of the glands, where the sporozoites invade preferentially. These results open up the possibilities of elucidating salivary gland-parasite interactions and generating transgenic mosquitoes refractory to parasites.Recent advances in the genetic engineering of anopheline mosquitoes have raised hopes for their use as … More new strategies for malaria control and provided a powerful tool to investigate mosquito-parasite interactions. Our research on transmission blockade using transgenic mosquitoes has been focused on the midgut stage of the parasites, interfering with ookinete invasion or oocyst differentiation. The molecule that we have introduced into a transgenic mosquito is CEL-III isolated from Cucumaria echinata (Marine invertebrate). CEL-III has strong hemolytic activity as well as haemaggrutination activity as a lectin. CEL-III gene expression was driven by the carboxypeptidase promoter, midgut specific promoter. As expected, CEL-III expression was induced in the midgut at 6h after a blood meal. We fed control and transgenic mosquitoes on the same infected rat and measure the numbers of oocysts formed. The transmission blocking efficacy of the transgenic mosquitoes was 91%, as compared with control mosquito. Thus, expression of CEL-III in the mosquito midgut severely reduced vector competency by inhibiting Plasmodium development. Because CEL-III targets to red blood cells but not the parasites, we anticipate that this CEL-III transgenic mosquito will be able to impair parasite development of all Plasmodium species. Less

疟疾孢子侵入蚊子的唾液腺，在唾液管中等待下一次吸血。唾液腺侵袭的过程和分子机制仍然是未知的。为了在斯氏按蚊中建立强大的唾液腺特异性转基因表达，我们获得了一个编码按蚊抗血小板聚集蛋白（AAPP）基因的唾液腺特异性启动子。aapp启动子是一种女性唾液腺特异性、血餐诱导的强启动子。利用这个启动子，我们产生了一个转基因An。在腺的远侧叶中表达丰富的Discosoma sp.红色荧光蛋白（DsRed），孢子子优先侵入。这些结果为阐明唾液腺与寄生虫的相互作用和产生对寄生虫不耐受的转基因蚊子提供了可能性。疟蚊基因工程的最新进展为它们作为疟疾控制的新策略提供了希望，并为研究蚊子与寄生虫的相互作用提供了有力的工具。我们利用转基因蚊子阻断传播的研究主要集中在寄生虫的中肠阶段，干扰卵母细胞入侵或卵囊分化。我们导入转基因蚊子的分子是从海生无脊椎动物黄瓜中分离出来的CEL-III。CEL-III作为凝集素具有较强的溶血活性和凝血活性。CEL-III基因的表达由羧基肽酶启动子、中肠特异性启动子驱动。正如预期的那样，在血餐后6小时，在中肠诱导了CEL-III的表达。我们将对照蚊子和转基因蚊子分别喂于同一只感染大鼠，并测量形成的卵囊数量。与对照蚊相比，转基因蚊的传播阻断率为91%。因此，CEL-III在蚊子中肠中的表达通过抑制疟原虫的发育严重降低了媒介能力。由于CEL-III靶向红细胞而不是寄生虫，我们预计这种CEL-III转基因蚊子将能够破坏所有疟原虫的寄生虫发育。少

项目成果

The development of vaccines against SARS corona virus in mice and SCID-PBL/hu mice.

• DOI: 10.1016/j.vaccine.2005.01.036
• 发表时间: 2005-03-18
• 期刊: Vaccine
• 影响因子: 5.5
• 作者: Okada M;Takemoto Y;Okuno Y;Hashimoto S;Yoshida S;Fukunaga Y;Tanaka T;Kita Y;Kuwayama S;Muraki Y;Kanamaru N;Takai H;Okada C;Sakaguchi Y;Furukawa I;Yamada K;Matsumoto M;Kase T;Demello DE;Peiris JS;Chen PJ;Yamamoto N;Yoshinaka Y;Nomura T;Ishida I;Morikawa S;Tashiro M;Sakatani M
• 通讯作者: Sakatani M

Novel recombinant BCG and DNA-vaccination against tuberculosis in a cynomolgus monkey model.

食蟹猴模型中针对结核病的新型重组 BCG 和 DNA 疫苗接种。

• 发表时间: 2005
• 期刊: Vaccine 23
• 作者: Ansari AA;Mayne AE;Onlamoon N;Pattanapanyasat K;Mori K;Villinger F.;大友一雄;大友一雄;大友 一雄;五島 敏芳;神立 孝一;岡田全司;岡田全司;岡田全司;岡田全司
• 通讯作者: 岡田全司

A novel alpha-proteobacterium invades the mitochondria of ovarian cells of the tick Ixodes ricinus.

一种新型α-变形菌侵入蓖麻蜱卵巢细胞的线粒体。

• 发表时间: 2004
• 期刊: Appl. Environ. Microbiol. 70
• 作者: Beninati;T.;N.Lo;L.Sacchi;C.Bandi;H.Noda;C.Genchi
• 通讯作者: C.Genchi

Wolbachia and nuclear-nuclear interactions contribute to reproductive incompatibility in the spider mite Panonychus mori (Acari : Tetranychidae). Tetranychidae)

沃尔巴克氏体和核-核相互作用导致红蜘蛛全爪螨（螨虫：叶螨科）的生殖不相容性。

• 发表时间: 2005
• 期刊: Heredity 94(2)
• 作者: Gotoh T;Noda H;Fujita T;Iwadate K;Higo Y;Saito S;Ohtsuka S.
• 通讯作者: Ohtsuka S.