Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Leonela Z. Carabajal Paladino; Ray Wilson; Priscilla Y. L. Tng; Vishaal Dhokiya; Elizabeth Keen; Piotr Cuber; Will Larner; Sara Rooney; Melanie Nicholls; Anastasia Uglow; Luke Williams; Michelle A. E. Anderson; Sanjay Basu; Philip T. Leftwich; Luke Alphey

doi:10.1101/2023.07.07.548128

Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Leonela Z. Carabajal Paladino, Ray Wilson, Priscilla Y. L. Tng, Vishaal Dhokiya, Elizabeth Keen, Piotr Cuber, Will Larner, Sara Rooney, Melanie Nicholls, Anastasia Uglow, Luke Williams, Michelle A. E. Anderson, Sanjay Basu, Philip T. Leftwich, Luke Alphey

Research output: Working paper › Preprint

Abstract

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. Here we present two improved methods for genetic manipulation in this species. 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.

Original language	English
Publisher	biorxiv
DOIs	https://doi.org/10.1101/2023.07.07.548128
Publication status	Published - 7 Jul 2023

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abstract = "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. Here we present two improved methods for genetic manipulation in this species. 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.",

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AU - Carabajal Paladino, Leonela Z.

AU - Wilson, Ray

AU - Tng, Priscilla Y. L.

AU - Dhokiya, Vishaal

AU - Keen, Elizabeth

AU - Cuber, Piotr

AU - Larner, Will

AU - Rooney, Sara

AU - Nicholls, Melanie

AU - Uglow, Anastasia

AU - Williams, Luke

AU - Anderson, Michelle A. E.

AU - Basu, Sanjay

AU - Leftwich, Philip T.

AU - Alphey, Luke

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AB - 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. Here we present two improved methods for genetic manipulation in this species. 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.

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Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Abstract

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