The mutagenic chain reaction: a method for autocatalyic gene dissemination

诱变链式反应：一种自催化基因传播的方法

• 批准号: 10614935
• 负责人: ETHAN BIER
• 金额: $ 32.39万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614935
• 关键词: Acceleration; Alleles; Anopheles Genus; Antibiotic Resistance; Antibodies; Antimalarials; Bacteria; Binding; Blastoderm; CRISPR gene drive; Child; Chromosomes; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Collaborations; Combination Drug Therapy; Complementary DNA; Complex; Culicidae; DNA; DNA Double Strand Break; DNA Repair; DNA cassette; Development; Disease; Drosophila genus; Drug resistance; Elements; Embryo; Event; Female; Foundations; Future; Genes; Genetic; Genome; Genomics; Germ Cells; Government; Grant; Guide RNA; Health; Heterozygote; Homologous Gene; Human; Image; Immunize; Individual; Inherited; Insect Vectors; Insecta; Insecticide Resistance; Insecticides; Intervention; Laboratories; Malaria; Mammals; Measures; Mediating; Methods; Mosaicism; Mus; Mutagenesis; Mutate; Parasite resistance; Parasites; Pathway interactions; Performance; Persons; Phlebotominae; Population; Poverty; Poverty Areas; Prevalence; Process; Prokaryotic Cells; Reaction; Reproduction; Resistance; Sand Flies; Sanitation; Scourge; Site; Source; Specific qualifier value; Sterility; System; Target Populations; Technology; Testing; Transgenes; Tsetse Flies; Update; Vector-transmitted infectious disease; Virus; Visualization; World Health Organization; blastomere structure; blocking factor; combat; cost; design; embryo cell; endonuclease; experimental study; flexibility; fly; food insecurity; functional restoration; gene drive system; genetic element; genetic technology; genetic variant; genomic locus; global health; imaging modality; loss of function; malaria transmission; mutagenic chain reaction; next generation; novel; pathogen; pragmatic implementation; prototype; rational design; repaired; tool; trait; transmission process; vector; welfare

Following a decade of significant strides forward, the global malaria eradication agenda has stalled, due in part to the accelerating emergence of insecticide-resistant mosquitoes and drug-resistant malarial parasites. The World Health Organization and others have called for the development of new strategies to help defeat this devastating disease that infects over 2 million people and killing over 400,000 annually, predominantly young children in impoverished regions. Gene-drives, which can bias inheritance of desired traits, offer a novel and promising strategy either to eliminate disease causing insect vectors, or to immunize them against pathogens. Such super-Mendelian CRIPSR-based gene-drive systems encode bipartite transgenic cassettes consisting of the Cas9 endonuclease and a guide RNA (gRNA), which directs DNA cleavage at the genomic site of insertion. In reproductive cells, such targeted cutting of the homologous chromosome results in copying the drive element at the cleavage site through homology directed repair, resulting in nearly all progeny inheriting the drive element and its cargo. My group has contributed to developing the first CRIPSR-based gene drive (or active genetic) systems in flies, mosquitoes, mammals, and bacteria. We also pioneered allelic-drive systems designed to bias inheritance of a favored allelic variant at a separate genetic locus. In addition, we have developed, and extensively tested, two types of self-copying drive neutralizing systems, both of which carry gRNAs, but no source of Cas9. ERACRs delete and replace gene-drives, while e-CHACRs copy themselves while mutating and inactivating the Cas9 transgene carried on a gene-drive. Small population cage experiments in flies and mosquitoes have shown that highly efficient gene-drives rapidly spread through target populations, and that ERACRs and e-CHACRs can reliably replace (ERACRs) or halt (e-CHACRs) a gene-drive element. In this grant, we propose first to develop a flexible two-component (split-drive or CHACR) system that can be genetically converted (or hacked) into a single full-drive system. The split and full drive elements are inserted into genes essential for viability or reproduction, and also carry recoded cDNAs of the targeted genes to restore function of those loci. These recoded systems benefit greatly from a phenomenon we discovered and refer to as lethal/sterile mosaicism, which dominantly eliminates loss-of-function alleles (mistakes) in the target gene generated by imprecise DNA repair events rather than the intended copying event. Next, we will develop and test next-generation ERACR and e-CHACRs able to eliminate or halt our recoded-drives, and also test a self-limiting drive system that slowly targets Cas9 for mutagenesis. In parallel to these drive experiments, we will delve into the mechanisms and timing of the drive process using a unique set of image-based genetic elements we have developed. We anticipate that the intellectual advances and implementable game-changing technologies from these studies will contribute importantly to solving critical global challenges in human health.

在取得了十年的重大进展之后，全球消灭疟疾议程停滞不前，原因是

项目成果

Development of an anti-Pfs230 monoclonal antibody as a Plasmodium falciparum gametocyte blocker.

开发抗 Pfs230 单克隆抗体作为恶性疟原虫配子体阻断剂。

• DOI: 10.21203/rs.3.rs-3757253/v1
• 发表时间: 2023
• 期刊: Research square
• 作者: Cuccurullo,EmiliaC;Dong,Yuemei;Simões,MariaL;Dimopoulos,George;Bier,Ethan
• 通讯作者: Bier,Ethan

Active genetics comes alive: Exploring the broad applications of CRISPR-based selfish genetic elements (or gene-drives): Exploring the broad applications of CRISPR-based selfish genetic elements (or gene-drives).

• DOI: 10.1002/bies.202100279
• 发表时间: 2022-08
• 期刊: BioEssays : news and reviews in molecular, cellular and developmental biology

Reversing insecticide resistance with allelic-drive in Drosophila melanogaster.

• DOI: 10.1038/s41467-021-27654-1
• 发表时间: 2022-01-12
• 期刊: Nature communications
• 影响因子: 16.6
• 作者: Kaduskar B;Kushwah RBS;Auradkar A;Guichard A;Li M;Bennett JB;Julio AHF;Marshall JM;Montell C;Bier E
• 通讯作者: Bier E

Genetic conversion of a split-drive into a full-drive element.

• DOI: 10.1038/s41467-022-35044-4
• 发表时间: 2023-01-12
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Terradas, Gerard;Bennett, Jared B.;Li, Zhiqian;Marshall, John M.;Bier, Ethan
• 通讯作者: Bier, Ethan

tgCRISPRi: efficient gene knock-down using truncated gRNAs and catalytically active Cas9.

• DOI: 10.1038/s41467-023-40836-3
• 发表时间: 2023-09-11
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Auradkar, Ankush;Guichard, Annabel;Kaduwal, Saluja;Sneider, Marketta;Bier, Ethan
• 通讯作者: Bier, Ethan