Highly specific chimeric DNA-RNA-guided genome editing with enhanced CRISPR-Cas12a system

• Main Authors: Hanseop Kim (Author), Wi-jae Lee (Author), Chan Hyoung Kim (Author), Yeounsun Oh (Author), Lee Wha Gwon (Author), Hyomin Lee (Author), Woojeung Song (Author), Junho K. Hur (Author), Kyung-Seob Lim (Author), Kang Jin Jeong (Author), Ki-Hoan Nam (Author), Young-Suk Won (Author), Kyeong-Ryoon Lee (Author), Youngjeon Lee (Author), Young-Hyun Kim (Author), Jae-Won Huh (Author), Bong-Hyun Jun (Author), Dong-Seok Lee (Author), Seung Hwan Lee (Author)
• Format: Book
• Published: Elsevier, 2022-06-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.

Summary

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a system is composed of a Cas12a effector that acts as a DNA-cleaving endonuclease and a crispr RNA (crRNA) that guides the effector to the target DNA. It is considered a key molecule for inducing target-specific gene editing in various living systems. Here, we improved the efficiency and specificity of the CRISPR-Cas12a system through protein and crRNA engineering. In particular, to optimize the CRISPR-Cas12a system at the molecular level, we used a chimeric DNA-RNA guide chemically similar to crRNA to maximize target sequence specificity. Compared with the wild-type (wt)-Cas12a system, when using enhanced Cas12a system (en-Cas12a), the efficiency and target specificity improved on average by 2.58 and 2.77 times, respectively. In our study, when the chimeric DNA-RNA-guided en-Cas12a effector was used, the gene-editing efficiency and accuracy were simultaneously increased. These findings could contribute to highly accurate genome editing, such as human gene therapy, in the near future.

Item Description

2162-2531
10.1016/j.omtn.2022.03.021