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Protocol for enhancing CRISPR-Cas9 genome editing using histone deacetylase inhibition and engineered virus-like
Mahbod Djamshidi1, Ryota Tanida1, Katayoun Heshmatzad2
1Robson DNA Sciences Centre, Arnie Charbonneau Cancer Institute, University of Calgary, Calgary, AB, Canada; Alberta Children's Hospital Research Institute, University of Calgary, Calgary, AB, Canada; Arthur JE Childs Comprehensive Cancer Institute, University of Calgary, Calgary, AB, Canada; Departments of Biochemistry and Molecular Biology, University of Calgary, Calgary, AB, Canada; Cumming School of Medicine, University of Calgary, Calgary, AB, Canada.
Abstract:
We present a 10-fold faster, accurate, and more efficient (FAME)-CRISPR-Cas9 gene editing workflow utilizing histone deacetylase inhibitor (HDACi)-mediated chromatin relaxation and engineered virus-like particle (eVLP) delivery of Cas9. We describe steps for optimizing HDACi concentration, euchromatinization timing, and Cas9 delivery/expression to improve CRISPR-Cas9 editing efficiency and efficacy. This protocol can eliminate the need for single-cell cloning and reduce experimental timelines up to 10-fold while minimizing HDACi-mediated toxicity. For complete details on the use and execution of this protocol, please refer to Djamshidi et al.1.
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