Next-generation gene drive for population modification of the malaria vector mosquito, Anopheles gambiae.
Next-generation gene drive for population modification of the malaria vector mosquito, Anopheles gambiae.
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DOI：
10.1073/pnas.2010214117
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发表时间：
2020-09-15
影响因子：
11.1
通讯作者：
James AA
中科院分区：
文献类型：
--
作者：
Carballar-Lejarazú R;Ogaugwu C;Tushar T;Kelsey A;Pham TB;Murphy J;Schmidt H;Lee Y;Lanzaro GC;James AA
控制媒介传播疾病传播的遗传系统正在从原理证明的发现阶段示范转向下一个发展阶段。成功的过渡需要满足目标产品简介中定义的安全性和有效性标准。我们在这里展示了一种基因工程疟疾蚊子载体冈比亚按蚊（Anopheles gambiae）的基于Cas9/向导rna的基因驱动组件，达到了功效和性能的关键目标产品谱要求。该系统旨在与编码抗寄生虫效应分子的基因结合，实现蚊子种群修饰，从而稳定和可持续地阻断疟疾寄生虫的传播。研究了一种基于Cas9/guide rna的基因驱动菌株AgNosCd-1，该菌株可将抗寄生虫效应分子传递给疟疾媒介冈比亚按蚊。驱动系统的目标是产生红眼表型的主要基因同源物。在小笼试验中，在一次释放基因驱动雄性后的6至10代内，可以观察到基因驱动雄性的完全引入。在试验中，没有由整合转基因引起的遗传负荷损害驱动性能。潜在的驱动抗性靶位等位基因出现的频率小于0.1，在定植和野生来源的非洲蚊子中，向导RNA靶位最普遍的5种多态性不能阻止Cas9/向导RNA复合物在体外切割。只有一个预测的脱靶位点在体外可切割，在体内观察到的缺失可以忽略不计。AgNosCd-1符合目标产品概况的关键性能标准，可以成为现场准备菌株的重要组成部分，用于改变蚊子种群以控制疟疾传播。
Genetic systems for controlling transmission of vector-borne diseases are moving from discovery-stage demonstrations of proofs-of-principle to the next phases of development. A successful transition requires meeting safety and efficacy criteria defined in target product profiles. We show here that the Cas9/guide RNA-based gene-drive components of a genetically-engineered malaria mosquito vector, Anopheles gambiae, achieve key target product profile requirements for efficacy and performance. This system is designed to achieve mosquito population modification when coupled with genes encoding antiparasite effector molecules and result in stable and sustainable blocking of malaria parasite transmission. A Cas9/guide RNA-based gene drive strain, AgNosCd-1, was developed to deliver antiparasite effector molecules to the malaria vector mosquito, Anopheles gambiae. The drive system targets the cardinal gene ortholog producing a red-eye phenotype. Drive can achieve 98 to 100% in both sexes and full introduction was observed in small cage trials within 6 to 10 generations following a single release of gene-drive males. No genetic load resulting from the integrated transgenes impaired drive performance in the trials. Potential drive-resistant target-site alleles arise at a frequency <0.1, and five of the most prevalent polymorphisms in the guide RNA target site in collections of colonized and wild-derived African mosquitoes do not prevent cleavage in vitro by the Cas9/guide RNA complex. Only one predicted off-target site is cleavable in vitro, with negligible deletions observed in vivo. AgNosCd-1 meets key performance criteria of a target product profile and can be a valuable component of a field-ready strain for mosquito population modification to control malaria transmission.
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影响因子：
3.7
作者：
Meredith JM;Underhill A;McArthur CC;Eggleston P
通讯作者：
Eggleston P
影响因子：
3.8
作者：
Carvalho DO;McKemey AR;Garziera L;Lacroix R;Donnelly CA;Alphey L;Malavasi A;Capurro ML
通讯作者：
Capurro ML
影响因子：
3.4
作者：
Carballar-Lejarazú R;James AA
通讯作者：
James AA
影响因子：
3.8
作者：
Macias VM;Jimenez AJ;Burini-Kojin B;Pledger D;Jasinskiene N;Phong CH;Chu K;Fazekas A;Martin K;Marinotti O;James AA
通讯作者：
James AA
影响因子：
2.1
作者：
James, Stephanie L.;Marshall, John M.;Nolan, Tony
通讯作者：
Nolan, Tony