Germline excision of transgenes in Aedes aegypti by homing endonucleases

Files

TR Number

Date

2013-04

Journal Title

Journal ISSN

Volume Title

Publisher

Nature Publishing Group

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.

Description

Keywords

engineered male mosquitos, yellow-fever mosquito, homologous, recombination, directed evolution, anopheles-gambiae, dna breaks, gene, drosophila, sequences, populations, multidisciplinary sciences

Citation

Aryan, A.; Anderson, M. A. E.; Myles, K. M.; Adelman, Z. N., "Germline excision of transgenes in Aedes aegypti by homing endonucleases," Scientific Reports 3:1603, (2013). DOI: 10.1038/srep01603.