Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti

Frontiers in Bioengineering and Biotechnology

Bioengineering and Biotechnology

Leonela Z. Carabajal Paladino¹  Michelle A. E. Anderson¹  Will Larner¹  Melanie Nicholls¹  Sara Rooney¹  Ray Wilson¹  Anastasia Uglow¹  Vishaal Dhokiya¹  Piotr Cuber¹  Philip T. Leftwich¹  Priscilla Y. L. Tng¹  Sanjay Basu¹  Luke Williams¹  Elizabeth Keen¹  Luke Alphey² 

[1] Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom;null;

关键词: integration;    plasmid;    cassette exchange;    transgenesis;    mosquito;   

DOI  :  10.3389/fbioe.2023.1254863

 received in 2023-07-07, accepted in 2023-08-31,  发布年份 2023

来源: Frontiers

【 摘 要 】

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.

【 授权许可】

Unknown   
Copyright © 2023 Carabajal Paladino, Wilson, Tng, Dhokiya, Keen, Cuber, Larner, Rooney, Nicholls, Uglow, Williams, Anderson, Basu, Leftwich and Alphey.