Grigoraki, Linda, Cowlishaw, Ruth, Nolan, Tony, Donnelly, Martin, Lycett, Gareth 
ORCID: 0000-0002-2422-053X and Ranson, Hilary
(2021)
CRISPR/Cas9 modified <i>An</i>. <i>gambiae</i> carrying <i>kdr</i> mutation L1014F functionally validate its contribution in insecticide resistance and combined effect with metabolic enzymes.
PLOS GENETICS, 17 (7).
e1009556-.
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CRISPRCas9 modified An. gambiae carrying kdr mutation L1014F functionally validate its contribution in insecticide resistanc.pdf - Open Access published version Download (2MB) | Preview |
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 |
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| Uncontrolled Keywords: | Animals, Animals, Genetically Modified, Anopheles, DDT, Pyrethrins, Permethrin, Nitriles, Piperonyl Butoxide, Glutathione Transferase, Insect Proteins, Fertility, Insecticide Resistance, Mutation, Female, Male, Genome, Insect, CRISPR-Cas Systems |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 06 Jul 2023 14:15 |
| Last Modified: | 19 Oct 2023 09:11 |
| DOI: | 10.1371/journal.pgen.1009556 |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3171469 |
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