Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Leonela Zusel Carabajal Paladino; Ray Wilson; Priscilla Tng; Vishaal Dhokiya; Elizabeth Keen; Piotr Cubr; Will Larner; Sara Rooney; Melanie Nicholls; Anastasia Uglow; Luke Williams; Michelle Anderson; Sanjay Basu; Philip Leftwich; Luke Stephen Alphey

doi:10.3389/fbioe.2023.1254863

Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Leonela Zusel Carabajal Paladino, Ray Wilson, Priscilla Tng, Vishaal Dhokiya, Elizabeth Keen, Piotr Cubr, Will Larner, Sara Rooney, Melanie Nicholls, Anastasia Uglow, Luke Williams, Michelle Anderson, Sanjay Basu, Philip Leftwich, Luke Stephen Alphey^*

^*Corresponding author for this work

Biology

Research output: Contribution to journal › Article › peer-review

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
Article number	1254863
Number of pages	12
Journal	Frontiers in Bioengineering and Biotechnology
Volume	11
DOIs	https://doi.org/10.3389/fbioe.2023.1254863
Publication status	Published - 22 Sept 2023

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10.3389/fbioe.2023.1254863Licence: CC BY

Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti
© 2023 Carabajal Paladino, Wilson, Tng, Dhokiya, Keen, Cuber, Larner, Rooney, Nicholls, Uglow, Williams, Anderson, Basu, Leftwich and Alphey.
Final published version, 2.01 MBLicence: CC BY

Safe Genes Daisy Drives
Alphey, L. S., Anderson, M. & Ang Xin De, J.
1/09/17 → 31/08/19
Project: Other project › Project from former institution

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Carabajal Paladino, L. Z., Wilson, R., Tng, P., Dhokiya, V., Keen, E., Cubr, P., Larner, W., Rooney, S., Nicholls, M., Uglow, A., Williams, L., Anderson, M., Basu, S., Leftwich, P., & Alphey, L. S. (2023). Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti. Frontiers in Bioengineering and Biotechnology, 11, Article 1254863. https://doi.org/10.3389/fbioe.2023.1254863

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abstract = "Genetic manipulation of Aedes aegypti is key to developing a deeper understanding of this insects{\textquoteright} 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 Zusel

AU - Wilson, Ray

AU - Tng, Priscilla

AU - Dhokiya, Vishaal

AU - Keen, Elizabeth

AU - Cubr, Piotr

AU - Larner, Will

AU - Rooney, Sara

AU - Nicholls, Melanie

AU - Uglow, Anastasia

AU - Williams, Luke

AU - Anderson, Michelle

AU - Basu, Sanjay

AU - Leftwich, Philip

AU - Alphey, Luke Stephen

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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.

U2 - 10.3389/fbioe.2023.1254863

DO - 10.3389/fbioe.2023.1254863

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Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Abstract

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Safe Genes Daisy Drives

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