Optimized genetic tools for neuroanatomical and functional mapping of the Aedes aegypti olfactory system

G3 Genes Genomes Genetics, Год журнала: 2025, Номер unknown

Опубликована: Янв. 24, 2025

Abstract The mosquito Aedes aegypti is an emerging model insect for invertebrate neurobiology. We detail the application of a dual transgenesis marker system that reports nature transgene integration with circular donor template CRISPR–Cas9-mediated homology-directed repair at target chemoreceptor genes. Employing this approach, we demonstrate establishment cell-type-specific T2A-QF2 driver lines A. olfactory co-receptor genes Ir8a and orco via canonical CO2 receptor complex gene Gr1 noncanonical involving duplication intended cassette separated by intervening plasmid sequence. Using Gr1+ sensory neurons as example, show introgression such QUAS responder transgenes into yellow cuticular pigmentation mutant strain facilitates transcuticular calcium imaging CO2-evoked neural activity on maxillary palps enhanced sensitivity relative to wild-type mosquitoes enveloped dark melanized cuticle. further apply Cre-loxP excision derive marker-free in-frame fusions clearly map axonal projection patterns from expressing these 3 chemoreceptors antennal lobe devoid background interference 3xP3-based fluorescent markers. clear recording responses in central brain region using genetically encoded indicators GCaMP6s CaMPARI2. Systematic optimized methods different stands enable mapping circuits peripheral levels coding high resolution.

Язык: Английский

Circadian rhythm of mosquitoes: Blood feeding and disease transmission

Decoding Infection and Transmission, Год журнала: 2025, Номер 3, С. 100050 - 100050

Опубликована: Янв. 1, 2025

Язык: Английский