This is a protocol preprint. Preprints are preliminary reports that have not undergone peer review. They should not be considered conclusive, used to inform clinical practice, or referenced by the media as validated information.
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 Creative Commons Attribution 4.0 International 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
Refer to the "attached file with the full procedure":https://www.nature.com/protocolexchange/system/uploads/5611/original/CIRCLE-seq_Supplementary_Protocol.pdf?1493740820
This is a list of supplementary files associated with this preprint. Click to download.
- CIRCLEseqSupplementaryProtocol.pdf
Supplementary document 1: CIRCLE-seq protocol CIRCLE-seq protocol
Version History
CURRENT STATUS: Posted
Version 1
Posted 08 Nov, 2017
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Subject Areas
Biotechnology
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)
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
Version History
CURRENT STATUS: Posted
Version 1
Posted 08 Nov, 2017
Metrics
Comments: 0
HTML Views: ...
Subject Areas
Biotechnology
License:
This work is licensed under a Creative Commons Attribution 4.0 International 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.
Refer to the "attached file with the full procedure":https://www.nature.com/protocolexchange/system/uploads/5611/original/CIRCLE-seq_Supplementary_Protocol.pdf?1493740820
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)