Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR
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 CRIS...
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2018-02-01
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doaj-4e8ebe0b64724308a7cd7c8cacdf92142021-07-02T01:44:17ZengOxford University PressG3: Genes, Genomes, Genetics2160-18362018-02-018265365810.1534/g3.117.113424Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPRMing LiOmar S. AkbariBradley J. WhiteAnopheles 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.http://g3journal.org/lookup/doi/10.1534/g3.117.1134Anophelesgene drivereverse geneticstransgenicsCRISPRCas9 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Ming Li Omar S. Akbari Bradley J. White |
spellingShingle |
Ming Li Omar S. Akbari Bradley J. White Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR G3: Genes, Genomes, Genetics Anopheles gene drive reverse genetics transgenics CRISPR Cas9 |
author_facet |
Ming Li Omar S. Akbari Bradley J. White |
author_sort |
Ming Li |
title |
Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR |
title_short |
Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR |
title_full |
Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR |
title_fullStr |
Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR |
title_full_unstemmed |
Highly Efficient Site-Specific Mutagenesis in Malaria Mosquitoes Using CRISPR |
title_sort |
highly efficient site-specific mutagenesis in malaria mosquitoes using crispr |
publisher |
Oxford University Press |
series |
G3: Genes, Genomes, Genetics |
issn |
2160-1836 |
publishDate |
2018-02-01 |
description |
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. |
topic |
Anopheles gene drive reverse genetics transgenics CRISPR Cas9 |
url |
http://g3journal.org/lookup/doi/10.1534/g3.117.1134 |
work_keys_str_mv |
AT mingli highlyefficientsitespecificmutagenesisinmalariamosquitoesusingcrispr AT omarsakbari highlyefficientsitespecificmutagenesisinmalariamosquitoesusingcrispr AT bradleyjwhite highlyefficientsitespecificmutagenesisinmalariamosquitoesusingcrispr |
_version_ |
1721344503900536832 |