Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti .

Autor:	Carabajal Paladino LZ; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Wilson R; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Tng PYL; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Dhokiya V; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Keen E; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Cuber P; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Larner W; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Rooney S; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Nicholls M; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Uglow A; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Williams L; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Anderson MAE; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Basu S; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Leftwich PT; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom., Alphey L; Arthropod Genetics Group, The Pirbright Institute, Woking, United Kingdom.
Jazyk:	angličtina
Zdroj:	Frontiers in bioengineering and biotechnology [Front Bioeng Biotechnol] 2023 Sep 22; Vol. 11, pp. 1254863. Date of Electronic Publication: 2023 Sep 22 (Print Publication: 2023).
DOI:	10.3389/fbioe.2023.1254863
Abstrakt:	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. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2023 Carabajal Paladino, Wilson, Tng, Dhokiya, Keen, Cuber, Larner, Rooney, Nicholls, Uglow, Williams, Anderson, Basu, Leftwich and Alphey.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu