Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Carabajal Paladino, Leonela Z., Wilson, Ray, Tng, Priscilla Y. L., Dhokiya, Vishaal, Keen, Elizabeth, Cuber, Piotr, Larner, Will, Rooney, Sara, Nicholls, Melanie, Uglow, Anastasia, Williams, Luke, Anderson, Michelle A. E., Basu, Sanjay, Leftwich, Philip T. ORCID: https://orcid.org/0000-0001-9500-6592 and Alphey, Luke (2023) Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti. Frontiers in Bioengineering and Biotechnology, 11. ISSN 2296-4185

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Abstract

Introduction: Genetic manipulation of Aedes aegypti is key to developing a deeper understanding of this insects’ biology, vector-virus interactions and makes future genetic control strategies possible. Despite some advances, this process remains laborious and requires highly skilled researchers and specialist equipment. Methods: Here we present two improved methods for genetic manipulation in this species. Use of transgenic lines which express Cre recombinase and a plasmid-based method for expressing PhiC31 when injected into early embryos. Results: Use of transgenic lines which express Cre recombinase allowed, by simple crossing schemes, germline or somatic recombination of transgenes, which could be utilized for numerous genetic manipulations. PhiC31 integrase based methods for site-specific integration of genetic elements was also improved, by developing a plasmid which expresses PhiC31 when injected into early embryos, eliminating the need to use costly and unstable mRNA as is the current standard. Discussion: Here we have expanded the toolbox for synthetic biology in Ae. aegypti. These methods can be easily transferred into other mosquito and even insect species by identifying appropriate promoter sequences. This advances the ability to manipulate these insects for fundamental studies, and for more applied approaches for pest control.

Item Type: Article
Faculty \ School: Faculty of Science > School of Biological Sciences
UEA Research Groups: Faculty of Science > Research Groups > Biosciences Teaching and Education Research
Depositing User: LivePure Connector
Date Deposited: 28 Nov 2023 03:01
Last Modified: 28 Nov 2023 03:01
URI: https://ueaeprints.uea.ac.uk/id/eprint/93839
DOI: 10.3389/fbioe.2023.1254863

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