title: Development of Light-Activated CRISPR Using Guide RNAs with Photocleavable Protectors.
abstract: The ability to remotely trigger CRISPR/Cas9 activity would enable new strategies to study cellular events with greater precision and complexity. In this work, we have developed a method to photocage the activity of the guide RNA called "CRISPR-plus" (CRISPR-precise light-mediated unveiling of sgRNAs). The photoactivation capability of our CRISPR-plus method is compatible with the simultaneous targeting of multiple DNA sequences and supports numerous modifications that can enable guide RNA labeling for use in imaging and mechanistic investigations.
authors.affiliation: Institute for Medical Engineering & Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
authors.forename: Piyush K
authors.identifier: 0000-0001-7153-442X
authors.initials: PK
authors.lastname: Jain
authors.affiliation: Institute for Medical Engineering & Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
authors.forename: Vyas
authors.initials: V
authors.lastname: Ramanan
authors.affiliation: Institute for Medical Engineering & Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
authors.forename: Arnout G
authors.identifier: 0000-0001-9346-344X
authors.initials: AG
authors.lastname: Schepers
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
authors.forename: Nisha S
authors.initials: NS
authors.lastname: Dalvie
authors.affiliation: Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
authors.forename: Apekshya
authors.initials: A
authors.lastname: Panda
authors.affiliation: Institute for Medical Engineering & Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
authors.forename: Heather E
authors.initials: HE
authors.lastname: Fleming
authors.affiliation: Institute for Medical Engineering & Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. sbhatia@mit.edu.|Department of Electrical Engineering and Computer Science, Marble Center for Cancer Nanomedicine, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. sbhatia@mit.edu.|Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA. sbhatia@mit.edu.|Broad Institute of MIT and Harvard, Cambridge, MA 02139, USA. sbhatia@mit.edu.|Howard Hughes Medical Institute, Cambridge, MA 02139, USA. sbhatia@mit.edu.
authors.forename: Sangeeta N
authors.initials: SN
authors.lastname: Bhatia
mesh_terms: D001483:Base Sequence; D064113:CRISPR-Cas Systems / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008027:Light; D009693:Nucleic Acid Hybridization; D010782:Photolysis / Q000528:radiation effects; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems; D049452:Green Fluorescent Proteins
keywords: CRISPR; DNA cleavage; gene technology; nucleic acids; photochemistry