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CRISPR-mediated knock-in of transgenes into the malaria vector Anopheles funestus

Quinn, Charlotte, Anthousi, Amalia, Wondji, Charles ORCID: and Nolan, Tony ORCID: (2021) 'CRISPR-mediated knock-in of transgenes into the malaria vector Anopheles funestus'. G3, Vol 11, Issue 8.

10.1093_g3journal_jkab201.pdf - Published Version
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The ability to introduce mutations, or transgenes, of choice to precise genomic locations has revolutionized our ability to understand howgenes and organisms work. In many mosquito species that are vectors of various human diseases, the advent of CRISPR genome editingtools has shed light on basic aspects of their biology that are relevant to their efficiency as disease vectors. This allows a better understand-ing of how current control tools work and opens up the possibility of novel genetic control approaches, such as gene drives, that deliber-ately introduce genetic traits into populations. Yet for theAnophelesfunestusmosquito, a significant vector of malaria in sub-SaharanAfrica and indeed the dominant vector species in many countries, transgenesis has yet to be achieved. We describe herein an optimizedtransformation system based on the germline delivery of CRISPR components that allows efficient cleavage of a previously validated geno-mic site and preferential repair of these cut sites via homology-directed repair (HDR), which allows the introduction of exogenous templatesequence, rather than end-joining repair. The rates of transformation achieved are sufficiently high that it should be able to introducealleles of choice to a target locus, and recover these, without the need to include additional dominant marker genes. Moreover, the highrates of HDR observed suggest that gene drives, which employ an HDR-type mechanism to ensure their proliferation in the genome, maybe well suited to work inA.funestus.

Item Type: Article
Subjects: QU Biochemistry > Genetics > QU 550 Genetic techniques. PCR. Chromosome mapping
QX Parasitology > Insects. Other Parasites > QX 510 Mosquitoes
QX Parasitology > Insects. Other Parasites > QX 515 Anopheles
WC Communicable Diseases > Tropical and Parasitic Diseases > WC 750 Malaria
Faculty: Department: Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI):
Depositing User: Stacy Murtagh
Date Deposited: 02 Aug 2021 08:58
Last Modified: 10 Dec 2021 12:19


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