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: 10624305
• 负责人: ETHAN BIER
• 金额: $ 46.26万
• 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/10624305
• 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; Endowment; 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; 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; transmission process; 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.

项目摘要 疟疾是目前全球影响最大的蚊媒疾病，使2.28亿人患病， 2018年有超过405，000人死亡，其中三分之二是幼儿-最脆弱的人口。几 按蚊属的蚊子种类可以作为引起疟疾的寄生虫的载体， 多年来，它们对杀虫剂的抗药性不断增加，阻碍了目前的控制方法，削弱了我们的反应。 最终的疾病爆发。全球化进一步使病媒和病原体自由移动， 在某些情况下，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， 代表着我们防治这一疾病的武器库中一个新的有希望的补充。这些技术 但在其发展过程中出现的问题很少。简而言之，基于 CRISPR由插入蚊子基因组中的Cas9和gRNA基因组成，所述基因位于 这种基因盒的排列赋予了它自我复制的特性， 它可以传播到野生型染色体上的相同位置。这种特性可以用来传播 在一个群体中，一个有益的性状将有助于减少疾病传播（群体改造），或 一种有害的特性，有助于减少蚊子数量（抑制）。 虽然这个过程是非常准确的，但它可能导致自传播的失败，并产生 靶基因座上的小突变阻止了基因驱动的进一步转化。这些“阻力” 在驱动过程中产生的“等位基因”已经被认为是 这些工具。此外，由于活性Cas9和gRNA在发育中的胚胎中的沉积， 当基因驱动从雌性遗传时，生物学允许这种抗性等位基因的广泛产生。 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， 野生按蚊种群的可靠工程。 为了使这些工具能够对全世界的疟疾发病率产生影响， 上述问题需要克服。为了解决这两个问题，在建议的三个目标中， 研究，我们将开发和优化果蝇中的三种平行技术， 随后将其应用于主要疟疾媒介斯氏按蚊。

项目成果

A nickase Cas9 gene-drive system promotes super-Mendelian inheritance in Drosophila.

• DOI: 10.1016/j.celrep.2022.110843
• 发表时间: 2022-05-24
• 期刊: CELL REPORTS
• 影响因子: 8.8
• 作者: Del Amo, Victor Lopez;Juste, Sara Sanz;Gantz, Valentino M.
• 通讯作者: Gantz, Valentino M.

Ethical Considerations for Gene Drive: Challenges of Balancing Inclusion, Power and Perspectives.

• DOI: 10.3389/fbioe.2022.826727
• 发表时间: 2022
• 期刊: Frontiers in bioengineering and biotechnology
• 影响因子: 5.7
• 作者: Kormos A;Lanzaro GC;Bier E;Santos V;Nazaré L;Pinto J;Aguiar Dos Santos A;James AA
• 通讯作者: James AA

Application of the Relationship-Based Model to Engagement for Field Trials of Genetically Engineered Malaria Vectors.

• DOI: 10.4269/ajtmh.20-0868
• 发表时间: 2020-12-21
• 期刊: The American journal of tropical medicine and hygiene
• 作者: Kormos A;Lanzaro GC;Bier E;Dimopoulos G;Marshall JM;Pinto J;Aguiar Dos Santos A;Bacar A;Sousa Pontes Sacramento Rompão H;James AA
• 通讯作者: James AA