Development of a gene drive system for locally confined and reversible population replacement in Aedes aegypti using chromosomal translocations

利用染色体易位开发埃及伊蚊局部受限和可逆种群替代的基因驱动系统

• 批准号: 277089463
• 负责人: Dr. Georg Oberhofer
• 金额: --
• 依托单位: Division of Biology and Biological Engineering
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/277089463?language=en
• 关键词: Development; gene; drive; system; locally

Aedes aegypti mosquitoes are the main vector for devastating diseases like dengue and yellow fever which are the cause for ten thousands of deaths each year, mainly among children. In the case of dengue fever the infection rates have approximately doubled every ten years since the 1970's, clearly showing that classic approaches using insecticides have failed to suppress the mosquito populations. Hence, novel strategies to control this global epidemic are in dire need.One approach to do this is with gene drive systems. They can be used to replace Aedes populations with strains that block the transmission of viruses. So far not a single gene drive system has been established in Aedes mosquitoes, the goal of the research proposed herein.The drive system I will build is based on chromosomal translocations. If translocation carrying mosquitoes mate into the wild, a proportion of the offspring is unviable due to chromosomal imbalance. This leads to a gradual loss of wildtype chromosomes and fixation of the translocation alleles. Among all the available drive systems, the translocation based drive was chosen because it is evolutionary stable, it does not spread to non-target areas and it can be reversed.To generate Aedes strains carrying translocations, two approaches will be pursued in parallel. First, translocation constructs will be integrated randomly into the Aedes genome with piggyBac transposase, followed by the activation of a linked homing endonuclease that induces double-stranded DNA breaks, which upon repair can form translocations. Second, translocations will be induced with the novel CRISPR/Cas9 system. Finally, the resulting strains will be evaluated in fitness assays and tested for their potential to drive population replacement.

埃及伊蚊是登革热和黄热病等毁灭性疾病的主要媒介，这些疾病每年导致数万人死亡，主要是儿童。在登革热的案例中，自20世纪70年代的S以来，感染率大约每十年翻一番，清楚地表明经典的杀虫剂方法未能抑制蚊子种群。因此，迫切需要新的策略来控制这一全球流行病。其中一种方法是利用基因驱动系统。它们可以用来用阻断病毒传播的菌株取代伊蚊种群。到目前为止，还没有在伊蚊中建立单一的基因驱动系统，本文提出的研究目标是：我将建立的驱动系统是基于染色体易位的。如果易位携带蚊子在野外交配，由于染色体不平衡，一定比例的后代无法存活。这会导致野生型染色体的逐渐丢失和易位等位基因的固定。在所有可用的驱动系统中，选择易位驱动是因为它进化稳定，不会扩散到非靶区，并且可以逆转。为了产生携带易位的伊蚊菌株，将并行使用两种方法。首先，易位结构将被随机整合到带有iggyBac转座酶的伊蚊基因组中，随后激活连接的归巢内切酶，导致双链DNA断裂，修复后可以形成易位。第二，新的CRISPR/Cas9系统将诱导易位。最后，将在适应性分析中对产生的菌株进行评估，并测试它们是否有可能推动种群更替。