Development of Light-Activated CRISPR Using Guide RNAs with Photocleavable Protectors

Piyush K Jain; Vyas Ramanan; Arnout G Schepers; Nisha S Dalvie; Apekshya Panda; Heather E Fleming; Sangeeta N Bhatia

doi:10.1002/anie.201606123

Development of Light-Activated CRISPR Using Guide RNAs with Photocleavable Protectors

Angew Chem Int Ed Engl. 2016 Sep 26;55(40):12440-4. doi: 10.1002/anie.201606123. Epub 2016 Aug 24.

Authors

Piyush K Jain¹, Vyas Ramanan¹, Arnout G Schepers¹, Nisha S Dalvie², Apekshya Panda², Heather E Fleming¹, Sangeeta N Bhatia^{3

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Affiliations

¹ Institute for Medical Engineering & Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
² Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
³ Institute for Medical Engineering & Science, Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. sbhatia@mit.edu.
⁴ Department of Electrical Engineering and Computer Science, Marble Center for Cancer Nanomedicine, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA. sbhatia@mit.edu.
⁵ Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA. sbhatia@mit.edu.
⁶ Broad Institute of MIT and Harvard, Cambridge, MA 02139, USA. sbhatia@mit.edu.
⁷ Howard Hughes Medical Institute, Cambridge, MA 02139, USA. sbhatia@mit.edu.

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.

Keywords: CRISPR; DNA cleavage; gene technology; nucleic acids; photochemistry.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Base Sequence
CRISPR-Cas Systems / genetics*
Green Fluorescent Proteins / genetics
HeLa Cells
Humans
Light
Nucleic Acid Hybridization
Photolysis / radiation effects
RNA, Guide, CRISPR-Cas Systems / chemistry
RNA, Guide, CRISPR-Cas Systems / metabolism*

Substances

RNA, Guide, CRISPR-Cas Systems
Green Fluorescent Proteins

Abstract

Publication types

MeSH terms

Substances

Grants and funding