Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR

G3: Genes, Genomes, Genetics

Ming Li^1¹ 

[1] Department of Entomology, University of California, Riverside, California 92521^1

关键词: Anopheles;    gene drive;    reverse genetics;    transgenics;    CRISPR;    Cas9;   

DOI  :  10.1534/g3.117.1134

学科分类：生物科学（综合）

来源: Genetics Society of America

【 摘 要 】

Anopheles mosquitoes transmit at least 200 million annual malaria infections worldwide. Despite considerable genomic resources, mechanistic understanding of biological processes in Anopheles has been hampered by a lack of tools for reverse genetics. Here, we report successful application of the CRISPR/Cas9 system for highly efficient, site-specific mutagenesis in the diverse malaria vectors Anopheles albimanus , A. coluzzii , and A. funestus . When guide RNAs (gRNAs) and Cas9 protein are injected at high concentration, germline mutations are common and usually biallelic, allowing for the rapid creation of stable mutant lines for reverse genetic analysis. Our protocol should enable researchers to dissect the molecular and cellular basis of anopheline traits critical to successful disease transmission, potentially exposing new targets for malaria control.

【 授权许可】

CC BY