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Nat Chem Biol ; 12(12): 1059-1064, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27723747

RESUMO

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.


Assuntos
Engenharia Genética/métodos , Integrases/genética , Optogenética , Recombinação Genética/genética , Animais , Células COS , Chlorocebus aethiops , Camundongos , Processos Fotoquímicos
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