Genome Editing Using CRISPR

Abstract

Genome editing is an indispensable technique utilized in life science and medicine. Development of genome editing tools with high efficiency and reliability has been a major challenge. Development of the CRISPR-Cas system for application in genome editing sparked a revolution due to its ease of use compared with previously developed genome editing tools. CRISPR nucleases are very versatile and fundamental genome editing tools, which recognize target sites in a guide RNA-dependent manner. CRISPR nucleases basically aim to generate DNA double-strand breaks (DSBs) at desired sites, but it was reported that CRISPR-mediated DNA DSBs frequently cause unforeseen large chromosomal deletions or genomic rearrangements and also induce the p53-mediated DNA damage response. Remarkably, through the development of new derivatives including base editor (BE) or prime editor (PE), the CRISPR-Cas system has the capability of overcoming such issues. Development of new genome editing tools based on the CRISPR-Cas system that are capable of enhancing the editing efficiency as well as compensating for deficiencies is ongoing. In this chapter, we report on the history of genome editing tools with an emphasis on the development of the CRISPR-Cas system and its status in the field of genome editing. In addition, we review the limitations of currently utilized genome editing tools that are based on the CRISPR-Cas system and consider the potential of the CRISPR-Cas system.