Frontiers in Bioengineering and Biotechnology | |
Optimizing CRE and PhiC31 mediated recombination in Aedes aegypti | |
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.
【 预 览 】
Files | Size | Format | View |
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RO202310129010984ZK.pdf | 2053KB | download |