Development of next-generation gene drive technologies for Anopheles population engineering

开发用于按蚊种群工程的下一代基因驱动技术

• 批准号: 10408862
• 负责人: ETHAN BIER
• 金额: $ 44.44万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10408862
• 关键词: Address; Affect; Alleles; Animals; Anopheles Genus; Antimalarials; Biology; Bypass; Child; Chromosomes; Clustered Regularly Interspaced Short Palindromic Repeats; Collaborations; Culicidae; DNA Double Strand Break; DNA cassette; Deposition; Development; Disease; Disease Outbreaks; Drosophila genus; Drosophila melanogaster; Effectiveness; Embryo; Embryonic Development; Endonuclease I; Engineering; Failure; Female; Future; Gene Conversion; Generations; Genes; Genetic Engineering; Genome; Goals; Guide RNA; Health; Immune; Impairment; Inherited; Insect Vectors; Insecta; Laboratories; Location; Malaria; Methods; Modification; Morbidity - disease rate; Mosquito-borne infectious disease; Mutation; Parasites; Pathway interactions; Persons; Pesticides; Population; Process; Production; Property; Public Health; Recycling; Research; Resistance; Technology; Testing; Translations; Work; base; burden of illness; design; disease transmission; egg; fighting; fitness; gene drive system; genetic technology; human pathogen; improved; insertion/deletion mutation; novel; offspring; pathogen; prevent; repaired; resistance allele; response; tool; trait; vector; vector mosquito

PROJECT SUMMARY Malaria is currently the most impactful mosquito-borne disease worldwide, sickening 228 million people and killing over 405,000 in 2018, 2/3 of which are young children — the most vulnerable demographic. Several mosquito species of the Anopheles genus can act as vectors of the parasite causing malaria, and in recent years their increasing resistance to pesticides is hampering current control methods and blunting our response to eventual disease outbreaks. Globalization is further allowing both vectors and pathogens to move freely and in certain situations to permanently establish themselves in new locations. CRISPR-based gene drive technologies for mosquito population engineering are being developed as they represent a new promising addition to our arsenal for fighting this disease. These technologies are up-and-coming, yet few issues have come up during their development. Briefly, a gene drive system based on CRISPR is composed of a Cas9 and a gRNA gene inserted in the mosquito genome at the location where the gRNA targets it. The arrangement of this genetic cassette endowed it with self-replicating properties that allow it to propagate to the same location on a wild-type chromosome. This property can be harnessed to spread within a population a beneficial trait that would help reducing disease transmission (population modification), or a deleterious trait to help reduce the mosquito population (suppression). While this process is extremely accurate, it can result in the failure of self-propagating, and the generation of small mutations at the targeted locus preventing further conversion by the gene drive. These “resistance alleles” generated during the drive process have been identified as a major hindrance to field applications of these tools. In addition, due to the deposition of active Cas9 and gRNA in the developing embryo, the mosquito biology allows an extensive production of such resistance alleles when a gene drive is inherited from a female. The long-term goal of this project is to develop powerful gene drive tools that can be used for the fast and reliable engineering of wild Anopheles populations. In order for these tools to be ready to have an impact on the malaria morbidity worldwide, the two issues described above need to be overcome. To tackle these two problems, in the three Aims of the proposed research, we will develop and optimize three parallel technologies in the fruit fly Drosophila melanogaster and subsequently apply them to the major malaria vector Anopheles stephensi.

项目总结 疟疾目前是全球影响最大的蚊媒疾病，使2.28亿人患病， 2018年死亡人数超过40.5万人，其中三分之二是幼儿--这是最脆弱的人群。几个 按蚊属的蚊子物种可以作为引起疟疾的寄生虫的媒介，在最近 多年来，它们对杀虫剂日益增长的抗药性阻碍了当前的控制方法，并削弱了我们的反应 最终的疾病暴发。全球化进一步允许病媒和病原体自由移动和 在某些情况下，在新的地点永久站稳脚跟。 基于CRISPR的蚊子种群工程基因驱动技术正在开发中，因为它们 代表着我们抗击这种疾病的新的有希望的补充。这些技术是 方兴未艾，但在发展过程中几乎没有出现什么问题。简而言之，一种基于 CRISPR由插入到蚊子基因组中的Cas9和gRNA基因组成，该基因位于 GRNA以它为靶标。这种遗传盒的排列赋予了它自我复制的特性，这使得 它会传播到野生型染色体上的相同位置。这一属性可以被利用来传播 在种群内有助于减少疾病传播的有益特征(种群修改)，或 这是一个有害的特征，有助于减少蚊子的数量(抑制)。 虽然这一过程非常准确，但它可能导致自传播失败，并生成 靶点上的微小突变阻止了基因驱动的进一步转化。这些“抵抗” 在驱赶过程中产生的等位基因已被确定为田间应用的主要障碍 这些工具。此外，由于活跃的Cas9和gRNA在发育中的胚胎中沉积，蚊子 生物学允许这样的抗性等位基因的广泛产生，当基因驱动是从女性遗传的时候。 该项目的长期目标是开发功能强大的基因驱动工具，可用于快速和 野生按蚊种群的可靠工程。 为了使这些工具能够对全球疟疾发病率产生影响，两个问题 上述问题需要克服。为了解决这两个问题，在提出的三个目标中 研究中，我们将开发和优化果蝇和黑腹果蝇的三项并行技术 随后将它们应用于主要的疟疾媒介斯氏按蚊。