Testing the causality between CYP9M10 and pyrethroid resistance using the TALEN and CRISPR/Cas9 technologies

Area of Science:

• Molecular Biology
• Entomology
• Genetics

Background:

• Genome editing advances enable gene function studies in non-model insects.
• Understanding insecticide resistance mechanisms is crucial for vector control.
• Overexpression of CYP9M10 is linked to pyrethroid resistance in Culex quinquefasciatus.

Purpose of the Study:

• To genetically verify the role of CYP9M10 in pyrethroid resistance using genome editing.
• To apply TALENs and CRISPR/Cas9 for targeted gene disruption in Culex quinquefasciatus.
• To develop an efficient method for TALE construction.

Main Methods:

• Utilized transcription activator-like effector nucleases (TALENs) and CRISPR/Cas9 systems.
• Developed a novel TALE construction approach using chemical cleavage.
• Induced frame-shifting mutations in the CYP9M10 gene in a resistant mosquito strain.

Main Results:

• Both TALENs and CRISPR/Cas9 successfully disrupted CYP9M10 in pyrethroid-resistant Culex quinquefasciatus.
• A mosquito line with a fully disrupted CYP9M10 haplotype exhibited over 100-fold reduction in larval pyrethroid resistance.
• Confirmed the causal relationship between CYP9M10 and pyrethroid resistance.

Conclusions:

• Genome editing provides a powerful tool for functional genomics in mosquitoes.
• CYP9M10 is a key determinant of pyrethroid resistance in Culex quinquefasciatus larvae.
• Targeting CYP9M10 could be a strategy for managing insecticide resistance.