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

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.