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

基本信息

批准号：
10278897
负责人：
ETHAN BIER
金额：
$ 44.43万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN DIEGO
依托单位国家：
美国
项目类别：
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/10278897
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 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

项目摘要

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.

项目概要疟疾是目前全球影响最大的蚊媒疾病，导致 2.28 亿人患病， 2018 年有超过 405,000 人死亡，其中 2/3 是幼儿——最脆弱的人群。按蚊属的蚊子可以作为引起疟疾的寄生虫的载体，并且最近多年来，它们对杀虫剂的耐药性不断增强，阻碍了当前的控制方法并削弱了我们的应对能力全球化进一步允许媒介和病原体自由流动。在某些情况下，他们可以在新的地点永久定居。用于蚊子种群工程的基于 CRISPR 的基因驱动技术正在开发中这些技术为我们抗击这种疾病的武器库增添了新的有前景的技术。简而言之，基于基因驱动系统，但在其开发过程中几乎没有出现任何问题。 CRISPR 由插入蚊子基因组中的 Cas9 和 gRNA 基因组成。 gRNA 以它为目标。这种基因盒的排列赋予了它自我复制的特性，使其能够进行自我复制。它可以传播到野生型染色体上的同一位置，可以利用这一特性进行传播。群体内的有益特征有助于减少疾病传播（群体改变），或有助于减少蚊子数量（抑制）的有害特征。虽然这个过程非常准确，但它可能会导致自传播失败，并产生目标位点的小突变阻止了基因驱动的进一步转化。驱动过程中产生的“等位基因”已被确定为现场应用的主要障碍此外，由于活性Cas9和gRNA在发育中的胚胎中沉积，蚊子。当基因驱动遗传自女性时，生物学允许大量产生此类抗性等位基因。该项目的长期目标是开发强大的基因驱动工具，可用于快速、准确地实现基因驱动。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，需要解决以下两个问题为了解决上述两个问题，提出了三个目标。研究，我们将在果蝇果蝇和果蝇中开发和优化三种并行技术随后将它们应用于主要疟疾媒介史氏按蚊。