title: A photoactivatable Cre-loxP recombination system for optogenetic genome engineering.
abstract: Genome engineering techniques represented by the Cre-loxP recombination system have been used extensively for biomedical research. However, powerful and useful techniques for genome engineering that have high spatiotemporal precision remain elusive. Here we develop a highly efficient photoactivatable Cre recombinase (PA-Cre) to optogenetically control genome engineering in vivo. PA-Cre is based on the reassembly of split Cre fragments by light-inducible dimerization of the Magnet system. PA-Cre enables sharp induction (up to 320-fold) of DNA recombination and is efficiently activated even by low-intensity illumination (∼0.04 W m-2) or short periods of pulsed illumination (∼30 s). We demonstrate that PA-Cre allows for efficient DNA recombination in an internal organ of living mice through noninvasive external illumination using a LED light source. The present PA-Cre provides a powerful tool to greatly facilitate optogenetic genome engineering in vivo.
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo, Japan.|Department of Rehabilitation and Regenerative Medicine, Columbia University, New York, USA.
authors.forename: Fuun
authors.initials: F
authors.lastname: Kawano
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo, Japan.
authors.forename: Risako
authors.initials: R
authors.lastname: Okazaki
authors.affiliation: Department of Rehabilitation and Regenerative Medicine, Columbia University, New York, USA.|Department of Pharmacology, Columbia University, New York, USA.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Yazawa
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo, Japan.
authors.forename: Moritoshi
authors.initials: M
authors.lastname: Sato
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D005818:Genetic Engineering / Q000379:methods*; D019426:Integrases / Q000235:genetics*; D051379:Mice; D062308:Optogenetics; D055668:Photochemical Processes; D011995:Recombination, Genetic / Q000235:genetics*
chemical_list: C045073:Cre recombinase; D019426:Integrases