CRISPR/Cas9 modified An. gambiae carrying kdr mutation L1014F functionally validate its contribution in insecticide resistance and combined effect with metabolic enzymes

Grigoraki, Linta ORCID: https://orcid.org/0000-0001-8997-0406, Cowlishaw, Ruth, Nolan, Tony ORCID: https://orcid.org/0000-0002-2982-8333, Donnelly, Martin ORCID: https://orcid.org/0000-0001-5218-1497, Lycett, Gareth ORCID: https://orcid.org/0000-0002-2422-053X and Ranson, Hilary ORCID: https://orcid.org/0000-0003-2332-8247 (2021) 'CRISPR/Cas9 modified An. gambiae carrying kdr mutation L1014F functionally validate its contribution in insecticide resistance and combined effect with metabolic enzymes'. PLoS Genetics, Vol 17, Issue 7, e1009556.

Text Grigoraki et al., 2021_manuscript - for library.docx - Accepted Version Available under License Creative Commons Attribution. Download (1MB)

Abstract

Insecticide resistance in Anopheles mosquitoes is a major obstacle in maintaining the momentum in reducing the malaria burden; mitigating strategies require improved understanding of the underlying mechanisms. Mutations in the target site of insecticides (the voltage gated sodium channel for the most widely used pyrethroid class) and over-expression of detoxification enzymes are commonly reported, but their relative contribution to phenotypic resistance remain poorly understood. Here we present a genome editing pipeline to introduce single nucleotide polymorphisms in An. gambiae which we have used to study the effect of the classical kdr mutation L1014F (L995F based on An. gambiae numbering), one of the most widely distributed resistance alleles. Introduction of 1014F in an otherwise fully susceptible genetic background increased levels of resistance to all tested pyrethroids and DDT ranging from 9.9-fold for permethrin to >24-fold for DDT. The introduction of the 1014F allele was sufficient to reduce mortality of mosquitoes after exposure to deltamethrin treated bednets, even as the only resistance mechanism present. When 1014F was combined with over-expression of glutathione transferase Gste2, resistance to permethrin increased further demonstrating the critical combined effect between target site resistance and detoxification enzymes in vivo. We also show that mosquitoes carrying the 1014F allele in homozygosity showed fitness disadvantages including increased mortality at the larval stage and a reduction in fecundity and adult longevity, which can have consequences for the strength of selection that will apply to this allele in the field.

Item Type:	Article
Subjects:	QX Parasitology > QX 4 General works QX Parasitology > Insects. Other Parasites > QX 510 Mosquitoes QX Parasitology > Insects. Other Parasites > QX 515 Anopheles QX Parasitology > Insects. Other Parasites > QX 600 Insect control. Tick control WA Public Health > Preventive Medicine > WA 240 Disinfection. Disinfestation. Pesticides (including diseases caused by)
Faculty: Department:	Biological Sciences > Vector Biology Department
Digital Object Identifer (DOI):	https://doi.org/10.1371/journal.pgen.1009556
Depositing User:	Samantha Sheldrake
Date Deposited:	08 Jul 2021 11:07
Last Modified:	08 Jul 2021 11:10
URI:	https://archive.lstmed.ac.uk/id/eprint/18137

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