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Target-Specific Precision of CRISPR-Mediated Genome Editing
Anob M Chakrabarti1, Tristan Henser-Brownhill2, Josep Monserrat2
1Bioinformatics and Computational Biology Laboratory, The Francis Crick Institute, 1 Midland Road, London NW1 1AT, UK; UCL Genetics Institute, Department of Genetics, Evolution and Environment, University College London, London WC1E 6BT, UK.
Molecular Cell
|December 18, 2018
Summary
CRISPR-Cas9 gene editing outcomes are unpredictable. This study reveals simple rules based on DNA sequences near the target site to predict editing precision and outcomes in human cells.
Area of Science:
- Molecular Biology
- Genetics
- Biotechnology
Background:
- The CRISPR-Cas9 system enables precise genome editing across organisms.
- Predicting specific editing outcomes at targeted genomic sites remains a challenge.
Purpose of the Study:
- To investigate indel profiles at over 1,000 genomic sites in human cells.
- To uncover general principles governing CRISPR-Cas9 mediated DNA editing outcomes.
- To identify predictive rules for editing precision and outcome.
Main Methods:
- Analysis of indel profiles at numerous genomic sites in human cells.
- Correlation analysis between editing precision, efficiency, and sequence features.
- Investigation of chromatin feature influence on indel profiles.
Main Results:
- Editing precision varies significantly across different genomic sites.
- Editing precision correlates with editing efficiency and preference for single-nucleotide insertions.
- Simple predictive rules, primarily dependent on the fourth nucleotide upstream of the PAM, were identified.
- Indel profiles are generally robust but can be modulated by chromatin features.
Conclusions:
- Predictable rules for CRISPR-Cas9 editing outcomes can be established based on local DNA sequence.
- Findings have implications for refining CRISPR technology for clinical applications.
- The study provides insights into DNA repair mechanisms, specifically broken end joining patterns.
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