title: Germline excision of transgenes in Aedes aegypti by homing endonucleases.
abstract: Aedes (Ae.) aegypti is the primary vector for dengue viruses (serotypes1-4) and chikungunya virus. Homing endonucleases (HEs) are ancient selfish elements that catalyze double-stranded DNA breaks (DSB) in a highly specific manner. In this report, we show that the HEs Y2-I-AniI, I-CreI and I-SceI are all capable of catalyzing the excision of genomic segments from the Ae. aegypti genome in a heritable manner. Y2-I-AniI demonstrated the highest efficiency at two independent genomic targets, with 20-40% of Y2-I-AniI-treated individuals producing offspring that had lost the target transgene. HE-induced DSBs were found to be repaired via the single-strand annealing (SSA) and non-homologous end-joining (NHEJ) pathways in a manner dependent on the availability of direct repeat sequences in the transgene. These results support the development of HE-based gene editing and gene drive strategies in Ae. aegypti, and confirm the utility of HEs in the manipulation and modification of transgenes in this important vector.
authors.affiliation: Fralin Life Science Institute and Department of Entomology, Virginia Tech, Blacksburg, VA 24061, USA.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Aryan
authors.forename: Michelle A E
authors.initials: MA
authors.lastname: Anderson
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Myles
authors.forename: Zach N
authors.initials: ZN
authors.lastname: Adelman
mesh_terms: D000330:Aedes / Q000235:genetics* / Q000821:virology*; D000818:Animals; D004260:DNA Repair / Q000235:genetics*; D019223:Densovirinae / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics*; D005854:Germ Cells / Q000502:physiology*; D019076:Transgenes / Q000235:genetics*
chemical_list: D004720:Endonucleases
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