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Method Article
Circularization for In vitro Reporting of Cleavage Effects (CIRCLE-seq)
Shengdar Tsai
Tsai Lab
Corresponding Author
J. Keith Joung
Massachusetts General Hospital and Harvard Medical School
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
CIRCLE-seq is a sensitive method for selective sequencing of Cas9-cleaved genomic DNA
molecules. Briefly, genomic DNA is circularized using a restriction-enzyme independent approach.
Successfully circularized molecules are enzymatically selected for by exonuclease treatment and
then treated with Cas9-gRNA complex. Only Cas9-linearized molecules can be subsequently
ligated to adapters for high-throughput sequencing.
Keywords
CIRCLE-seq
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- supplement0.pdf
Supplementary document 1: CIRCLE-seq protocol CIRCLE-seq protocol
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Associated Publications
CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR–Cas9 nuclease off-targets
by Shengdar Q Tsai, Nhu T Nguyen, Jose Malagon-Lopez, +3
Nature Methods (02 May, 2017)
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This is a preprint. It has not completed peer review.
Method Article
Circularization for In vitro Reporting of Cleavage Effects (CIRCLE-seq)
Shengdar Tsai
Tsai Lab
Corresponding Author
J. Keith Joung
Massachusetts General Hospital and Harvard Medical School
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 08 Nov, 2017
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biotechnology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
CIRCLE-seq is a sensitive method for selective sequencing of Cas9-cleaved genomic DNA
molecules. Briefly, genomic DNA is circularized using a restriction-enzyme independent approach.
Successfully circularized molecules are enzymatically selected for by exonuclease treatment and
then treated with Cas9-gRNA complex. Only Cas9-linearized molecules can be subsequently
ligated to adapters for high-throughput sequencing.
Associated Publications
CIRCLE-seq: a highly sensitive in vitro screen for genome-wide CRISPR–Cas9 nuclease off-targets
by Shengdar Q Tsai, Nhu T Nguyen, Jose Malagon-Lopez, +3
Nature Methods (02 May, 2017)