Engineered resistance to Plasmodium falciparum development in transgenic Anopheles stephensi.
Engineered resistance to Plasmodium falciparum development in transgenic Anopheles stephensi.
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DOI：
10.1371/journal.ppat.1002017
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发表时间：
2011-04
期刊：
影响因子：
6.7
通讯作者：
James AA
中科院分区：
文献类型：
--
作者：
Isaacs AT;Li F;Jasinskiene N;Chen X;Nirmala X;Marinotti O;Vinetz JM;James AA
转座子介导的转化用于产生斯氏按蚊，其表达设计用于靶向人类疟疾寄生虫恶性疟原虫的单链抗体（scFv）。scFv m1C3、m4B7和m2A10来源于小鼠单克隆抗体，其抑制中肠的动合子侵入或唾液腺的子孢子侵入。靶向寄生虫表面的scFv m4B7和m2A10与冈比亚按蚊抗微生物肽Cecropin A融合。将先前表征的按蚊顺式作用DNA调控元件包括在转基因中以协调scFv产生与寄生虫发育。基因扩增和免疫印迹分析表明，在血液喂养的女性转基因表达的启动子特异性增加。当用恶性疟原虫攻击时，表达每种scFv基因的转基因蚊子品系具有比对照显著更低的感染水平。消灭疟疾将需要病媒控制战略，这种战略既能自我维持，又不受受感染的人和蚊子迁移的影响。用抗寄生虫发育的转基因品系取代易感染疟疾的野生蚊子种群，可以中断疾病传播周期，支持根除工作。生产抗恶性疟原虫的蚊子是研究种群替代战略的必要的第一步。在这里，我们展示了一个。stephensi工程化以产生恶性疟原虫靶向效应分子，对这种重要的人类疟疾寄生虫具有抗性。三种效应分子中的两种代表了来自蚊子和小鼠免疫系统的成分的新组合。这些分子的一个重要特征是它们不太可能显著伤害蚊子，因为蚊子组分是具有抗疟原虫活性的按蚊抗微生物肽，而另一种组分是基于鼠抗体，所述鼠抗体是根据其特异性结合寄生虫蛋白的能力而选择的。具有这种设计的转基因加上基因驱动系统可以与疫苗和药物一起使用，作为根除疟疾长期战略的一部分，在当地可持续地消灭疟疾。
Transposon-mediated transformation was used to produce Anopheles stephensi that express single-chain antibodies (scFvs) designed to target the human malaria parasite, Plasmodium falciparum. The scFvs, m1C3, m4B7, and m2A10, are derived from mouse monoclonal antibodies that inhibit either ookinete invasion of the midgut or sporozoite invasion of salivary glands. The scFvs that target the parasite surface, m4B7 and m2A10, were fused to an Anopheles gambiae antimicrobial peptide, Cecropin A. Previously-characterized Anopheles cis-acting DNA regulatory elements were included in the transgenes to coordinate scFv production with parasite development. Gene amplification and immunoblot analyses showed promoter-specific increases in transgene expression in blood-fed females. Transgenic mosquito lines expressing each of the scFv genes had significantly lower infection levels than controls when challenged with P. falciparum. Malaria eradication will require vector-control strategies that are both self-sustaining and not affected by migration of infected humans and mosquitoes. Replacement of wild malaria-susceptible mosquito populations with transgenic strains refractory to parasite development could interrupt the cycle of disease transmission and support eradication efforts. Production of P. falciparum-resistant mosquitoes is a necessary first step towards investigating the population replacement strategy. Here we show that An. stephensi engineered to produce P. falciparum-targeting effector molecules are resistant to this important human malaria parasite. Two of the three effector molecules represent a novel combination of components derived from the immune systems of mosquitoes and mice. An important feature of these molecules is that they are unlikely to significantly harm the mosquito, as the mosquito component is an Anopheles antimicrobial peptide with activity against Plasmodium, while the other component is based on a murine antibody selected for its ability to bind specifically to a parasite protein. Transgenes with this design coupled with a gene-drive system could be used alongside vaccines and drugs to provide sustainable local elimination of malaria as part of a long-term strategy for eradication.
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DOI：
10.1084/jem.174.5.1203
发表时间：
1991-11-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Barr PJ;Green KM;Gibson HL;Bathurst IC;Quakyi IA;Kaslow DC
通讯作者：
Kaslow DC
影响因子：
5.4
作者：
ANKER, R;ZAVALA, F;POLLOK, BA
通讯作者：
POLLOK, BA
影响因子：
2.4
作者：
Horn, C;Wimmer, EA
通讯作者：
Wimmer, EA
影响因子：
15.8
作者：
Marshall, John M.;Taylor, Charles E.
通讯作者：
Taylor, Charles E.
影响因子：
6.7
作者：
Corby-Harris V;Drexler A;Watkins de Jong L;Antonova Y;Pakpour N;Ziegler R;Ramberg F;Lewis EE;Brown JM;Luckhart S;Riehle MA
通讯作者：
Riehle MA