PT  - JOURNAL ARTICLE
AU  - Ming Li
AU  - Omar S. Akbari
AU  - Bradley J. White
TI  - Highly efficient site-specific mutagenesis in Malaria mosquitoes using CRISPR
AID  - 10.1101/157867
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 157867
4099  - http://biorxiv.org/content/early/2017/06/30/157867.short
4100  - http://biorxiv.org/content/early/2017/06/30/157867.full
AB  - 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, Anopheles coluzzii, and Anopheles funestus. When guide RNAs and Cas9 are injected at high concentration, germline mutations are common and usually bi-allelic 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.