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Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

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...

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Autores principales: 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., Alphey, Luke
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10557486/
https://www.ncbi.nlm.nih.gov/pubmed/37811374
http://dx.doi.org/10.3389/fbioe.2023.1254863
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author 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.
Alphey, Luke
author_facet 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.
Alphey, Luke
author_sort Carabajal Paladino, Leonela Z.
collection PubMed
description 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.
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spelling pubmed-105574862023-10-07 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. Alphey, Luke Front Bioeng Biotechnol Bioengineering and Biotechnology 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. Frontiers Media S.A. 2023-09-22 /pmc/articles/PMC10557486/ /pubmed/37811374 http://dx.doi.org/10.3389/fbioe.2023.1254863 Text en Copyright © 2023 Carabajal Paladino, Wilson, Tng, Dhokiya, Keen, Cuber, Larner, Rooney, Nicholls, Uglow, Williams, Anderson, Basu, Leftwich and Alphey. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
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.
Alphey, Luke
Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti
title Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti
title_full Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti
title_fullStr Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti
title_full_unstemmed Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti
title_short Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti
title_sort optimizing cre and phic31 mediated recombination in aedes aegypti
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10557486/
https://www.ncbi.nlm.nih.gov/pubmed/37811374
http://dx.doi.org/10.3389/fbioe.2023.1254863
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