Photoactivatable Cre recombinase 3.0 for in vivo mouse applications

Kumi Morikawa, Kazuhiro Furuhashi, Carmen Sena-Tomas, Alvaro L. Garcia-Garcia, Ramsey Bekdash, Alison D. Klein, Nicholas Gallerani, Hannah E. Yamamoto, Seon-Hye E. Park, Grant S. Collins, Fuun Kawano, Moritoshi Sato, Chyuan-Sheng Lin, Kimara L. Targoff, Edmund Au, Michael C. Salling and Masayuki Yazawa ()
Additional contact information
Kumi Morikawa: Columbia University
Kazuhiro Furuhashi: Columbia University
Carmen Sena-Tomas: Columbia University
Alvaro L. Garcia-Garcia: Columbia University
Ramsey Bekdash: Columbia University
Alison D. Klein: Columbia University
Nicholas Gallerani: Columbia University
Hannah E. Yamamoto: Columbia University
Seon-Hye E. Park: Columbia University
Grant S. Collins: Louisiana State University Health Sciences Center
Fuun Kawano: Columbia University
Moritoshi Sato: The University of Tokyo
Chyuan-Sheng Lin: Columbia University
Kimara L. Targoff: Columbia University
Edmund Au: Columbia University
Michael C. Salling: Louisiana State University Health Sciences Center
Masayuki Yazawa: Columbia University

Nature Communications, 2020, vol. 11, issue 1, 1-11

Abstract: Abstract 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.

Date: 2020
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DOI: 10.1038/s41467-020-16030-0

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