Genetics tools for corpora allata specific gene expression in Aedes aegypti mosquitoes

• Published in: Scientific reports Vol. 12; no. 1; p. 20426
• Main Authors: Nouzova, Marcela, Edwards, Marten J., DeGennaro, Matthew, Leyva, Dennys, Tose, Lilian V., Fernandez-Lima, Francisco, Noriega, Fernando G.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 28-11-2022; Nature Publishing Group; Nature Portfolio
• Subjects: 631/1647/1511; 631/601/1466; Aedes - genetics; Aedes aegypti; Animals; Animals, Genetically Modified; Aquatic insects; Corpora Allata; CRISPR; Culicidae; Drosophila; Fluorescent indicators; Gene Expression; Genetics; Green fluorescent protein; Green Fluorescent Proteins - genetics; Humanities and Social Sciences; Juvenile Hormones; Mosquitoes; multidisciplinary; Promoters; Science; Science (multidisciplinary); Transcription initiation; Transgenes
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-022-25009-4

Juvenile hormone (JH) is synthesized by the
corpora allata
(CA) and controls development and reproduction in insects. Therefore, achieving tissue-specific expression of transgenes in the CA would be beneficial for mosquito research and control. Different CA promoters have been used to drive transgene expression in
Drosophila
, but mosquito CA-specific promoters have not been identified. Using the CRISPR/Cas9 system, we integrated transgenes encoding the reporter green fluorescent protein (GFP) close to the transcription start site of juvenile hormone acid methyl transferase (JHAMT), a locus encoding a JH biosynthetic enzyme, specifically and highly expressed in the CA of
Aedes aegypti
mosquitoes. Transgenic individuals showed specific GFP expression in the CA but failed to reproduce the full pattern of
jhamt
spatiotemporal expression. In addition, we created GeneSwitch driver and responder mosquito lines expressing an inducible fluorescent marker, enabling the temporal regulation of the transgene via the presence or absence of an inducer drug. The use of the GeneSwitch system has not previously been reported in mosquitoes and provides a new inducible binary system that can control transgene expression in
Aedes aegypti
.