title: Photoactivatable CRISPR-Cas9 for optogenetic genome editing.
abstract: We describe an engineered photoactivatable Cas9 (paCas9) that enables optogenetic control of CRISPR-Cas9 genome editing in human cells. paCas9 consists of split Cas9 fragments and photoinducible dimerization domains named Magnets. In response to blue light irradiation, paCas9 expressed in human embryonic kidney 293T cells induces targeted genome sequence modifications through both nonhomologous end joining and homology-directed repair pathways. Genome editing activity can be switched off simply by extinguishing the light. We also demonstrate activation of paCas9 in spatial patterns determined by the sites of irradiation. Optogenetic control of targeted genome editing should facilitate improved understanding of complex gene networks and could prove useful in biomedical applications.
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo, Japan.
authors.forename: Yuta
authors.initials: Y
authors.lastname: Nihongaki
authors.affiliation: Graduate School of Arts and Sciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo, Japan.
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: Takahiro
authors.initials: T
authors.lastname: Nakajima
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: D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics* / Q000528:radiation effects*; D005818:Genetic Engineering / Q000379:methods*; D057809:HEK293 Cells; D006801:Humans; D008957:Models, Genetic; D008969:Molecular Sequence Data; D062308:Optogenetics / Q000379:methods*; D034622:RNA Interference