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Development of Light‐Activated CRISPR Using Guide RNAs with Photocleavable Protectors
Author(s) -
Jain Piyush K.,
Ramanan Vyas,
Schepers Arnout G.,
Dalvie Nisha S.,
Panda Apekshya,
Fleming Heather E.,
Bhatia Sangeeta N.
Publication year - 2016
Publication title -
angewandte chemie
Language(s) - English
Resource type - Journals
eISSN - 1521-3757
pISSN - 0044-8249
DOI - 10.1002/ange.201606123
Subject(s) - crispr , cas9 , rna , guide rna , computational biology , dna , computer science , chemistry , nanotechnology , biology , biochemistry , gene , materials science
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.