Title:
METHOD FOR DETECTING OFF-TARGET SITE OF GENETIC SCISSORS IN GENOME
Document Type and Number:
WIPO Patent Application WO/2016/076672
Kind Code:
A1
Abstract:
The present invention relates to a method for detecting an off-target site of genetic scissors in the genome and, specifically, to a method for detecting an off-target site through data analysis by subjecting the genome isolated in vitro to genetic scissors to cut the genome and then performing whole genome sequencing of the genome (Digenome-seq), and to a method for selecting a target site of RGEN, which minimizes the off-target effect, using the detection method. The Digenome-seq of the present invention can detect the off-target site of the genetic scissors in the genome level at a high degree of reproducibility, and thus can be used in the manufacture of genetic scissors having high target specificity and the study of the genetic scissors.
Inventors:
KIM JIN SOO (KR)
KIM DAE SIK (KR)
BAE SANG SU (KR)
KIM DAE SIK (KR)
BAE SANG SU (KR)
Application Number:
PCT/KR2015/012255
Publication Date:
May 19, 2016
Filing Date:
November 13, 2015
Export Citation:
Assignee:
INST BASIC SCIENCE (KR)
International Classes:
C12Q1/68; C12N9/22; G16B30/10
Foreign References:
| US20140295556A1 | 2014-10-02 |
Other References:
CHO ET AL.: "Analysis of Off-Target Effects of CRISPR/Cas-derived RNA-Guided Endonucleases and Nickases", GENOME RESEARCH, vol. 24, January 2014 (2014-01-01), pages 132 - 141, XP055227885
KUSCU ET AL.: "Genome-Wide Analysis Reveals Characteristics of Off-Target Sites Bound by the Cas9 Endonuclease", NATURE BIOTECHNOLOGY, vol. 32, no. 7, July 2014 (2014-07-01), pages 677 - 683, XP055382577
PATTANAYAK ET AL.: "High-Throughput Profiling of Off-Target DNA Cleavage Reveals RNA-Programmed Cas9 Nuclease Specificity", NATURE BIOTECHNOLOGY, vol. 31, no. 9, 2013, pages 839 - 843, XP055294934
SHEN ET AL.: "Efficient Genome Modification by CRISPR-Cas9 Nickase with Minimal Off-Target Effects", NATURE METHODS, vol. 11, no. 4, April 2014 (2014-04-01), pages 399 - 402, XP055227888
KIM ET AL.: "Digenome-Seq: Genome-Wide Profiling of CRISPR-Cas9 Off-Target Effects in Human Cells", NATURE METHODS, vol. 12, no. 3, March 2015 (2015-03-01), pages 237 - 243, XP055287797
See also references of EP 3219810A4
KUSCU ET AL.: "Genome-Wide Analysis Reveals Characteristics of Off-Target Sites Bound by the Cas9 Endonuclease", NATURE BIOTECHNOLOGY, vol. 32, no. 7, July 2014 (2014-07-01), pages 677 - 683, XP055382577
PATTANAYAK ET AL.: "High-Throughput Profiling of Off-Target DNA Cleavage Reveals RNA-Programmed Cas9 Nuclease Specificity", NATURE BIOTECHNOLOGY, vol. 31, no. 9, 2013, pages 839 - 843, XP055294934
SHEN ET AL.: "Efficient Genome Modification by CRISPR-Cas9 Nickase with Minimal Off-Target Effects", NATURE METHODS, vol. 11, no. 4, April 2014 (2014-04-01), pages 399 - 402, XP055227888
KIM ET AL.: "Digenome-Seq: Genome-Wide Profiling of CRISPR-Cas9 Off-Target Effects in Human Cells", NATURE METHODS, vol. 12, no. 3, March 2015 (2015-03-01), pages 237 - 243, XP055287797
See also references of EP 3219810A4
Attorney, Agent or Firm:
SON, MIN (KR)
손민 (KR)
손민 (KR)
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