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Photoactivatable Cre recombinase 3.0 for in vivo mouse applications
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Photoactivatable Cre recombinase 3.0 for in vivo mouse applications
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Journal Article
Photoactivatable Cre recombinase 3.0 for in vivo mouse applications
2020
Overview
Optogenetic genome engineering tools enable spatiotemporal control of gene expression and provide new insight into biological function. Here, we report the new version of genetically encoded photoactivatable (PA) Cre recombinase, PA-Cre 3.0. To improve PA-Cre technology, we compare light-dimerization tools and optimize for mammalian expression using a CAG promoter, Magnets, and 2A self-cleaving peptide. To prevent background recombination caused by the high sequence similarity in the dimerization domains, we modify the codons for mouse gene targeting and viral production. Overall, these modifications significantly reduce dark leak activity and improve blue-light induction developing our new version, PA-Cre 3.0. As a resource, we have generated and validated AAV-PA-Cre 3.0 as well as two mouse lines that can conditionally express PA-Cre 3.0. Together these new tools will facilitate further biological and biomedical research.
Previous versions of photoactivatable Cre recombinase (PA-Cre) suffered from unintentional recombination in dark conditions. Here, the authors develop an improved version of PA-Cre, called PA-Cre 3.0, which shows reduced leakiness and improved efficiency upon activation, and make mouse lines that express PA-Cre 3.0 conditionally.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 38
/ 42/41
/ 64/60
/ 82/80
/ 96/44
/ Animals
/ Codons
/ Genetic Engineering - methods
/ Genetics
/ Genomes
/ Humanities and Social Sciences
/ Light
/ Magnets
/ Male
/ Mice
/ Promoter Regions, Genetic - genetics
/ Promoter Regions, Genetic - radiation effects
/ Recombination, Genetic - genetics
/ Recombination, Genetic - radiation effects
/ Science
