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Characterization of distinct mutation patterns by CRISPR-Cas9 and CRISPR-Cpf1 genome editing systems

Molecular & Cellular Toxicology 2019년 15권 4호 p.383 ~ 389
 ( Bae Tae-Geun ) - Kyung Hee University Graduate School Department of Medicine

 ( Hwang Woo-Chang ) - Seoul National University Research Institute of Basic Sciences
 ( Lee Do-Hyeon ) - Pusan National University School of Computer Science and Engineering
 ( Song Gil-Tae ) - Pusan National University School of Computer Science and Engineering
 ( Hur Jun-Seok W. ) - Korea University College of Medicine Department of Neurosurgery
 ( Hur Jun-Ho K. ) - Kyung Hee University College of Medicine Department of Pathology

Abstract


Backgrounds: The differences of DNA mutation patterns of genome editing by CRISPR (Clustered regularly interspaced Short Palindromic Repeats)-Cas9 and CRISPR-Cpf1 is not completely understood.

Methods: CRISPR-Cas9 or CRISPR-Cpf1 is applied for genome editing at the exact same four genomic locations in human embryonic kidney 293T (HEK293T) cells. The mutation patterns of the target genomic loci were analyzed by targeted deep sequencing.

Results: The mutation patterns of CRISPR-Cas9 and CRISPR-Cpf1 showed prominent differences. CRISPR-Cpf1 mediated genome engineering almost always resulted in deletion, while CRISPR-Cas9 showed remarkably high rate of insertion mutation of 1 base pair. The deletion patterns of CRISPR-Cas9 and CRISPR-Cpf1 were also different. The deletion patterns of CRISPR-Cas9 were composed of diverse and evenly distributed deletion patterns, while the mutation patterns of CRISPR-Cpf1 were composed of fewer major patterns.

Conclusion: Genome editing by CRISPR-Cas9 and CRISPR-Cpf1 shows different characteristics.

키워드

Genome editing; CRISPR; Cas9; Cpf1; Mutation patterns
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