Abstract

Although prime editing is a promising genome editing method, the efficiency of prime editor 2 (PE2) is often insufficient. Here we generate a more efficient variant of PE2, named hyPE2, by adding the Rad51 DNA-binding domain. When tested at endogenous sites, hyPE2 shows a median of 1.5- or 1.4- fold (range, 0.99- to 2.6-fold) higher efficiencies than PE2; furthermore, at sites where PE2-induced prime editing is very inefficient (efficiency < 1%), hyPE2 enables prime editing with efficiencies ranging from 1.1% to 2.9% at up to 34% of target sequences, potentially facilitating prime editing applications.

While prime editing is a promising technology, PE2 systems often have low efficiency. Here the authors fuse a Rad51 DNA-binding domain to create hyPE2 with improved editing efficiency.

Details

Title
Generation of a more efficient prime editor 2 by addition of the Rad51 DNA-binding domain
Author
Song Myungjae 1 ; Lim, Jung Min 2   VIAFID ORCID Logo  ; Seonwoo, Min 3 ; Jeong-Seok, Oh 4   VIAFID ORCID Logo  ; Kim Dong Young 1 ; Jae-Sung, Woo 4 ; Nishimasu Hiroshi 5   VIAFID ORCID Logo  ; Sung-Rae, Cho 6 ; Yoon Sungroh 7   VIAFID ORCID Logo  ; Kim Hyongbum Henry 8   VIAFID ORCID Logo 

1  Yonsei University College of Medicine, Department of Pharmacology, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Yonsei University College of Medicine, Graduate School of Medical Science, Brain Korea 21 Plus Project for Medical Sciences, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454) 
2  Yonsei University College of Medicine, Department of Pharmacology, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454) 
3  Seoul National University, Department of Electrical and Computer Engineering, Seoul, Republic of Korea (GRID:grid.31501.36) (ISNI:0000 0004 0470 5905) 
4  Korea University, Department of Life Sciences, Seoul, Republic of Korea (GRID:grid.222754.4) (ISNI:0000 0001 0840 2678) 
5  The University of Tokyo, Structural Biology Division, Research Center for Advanced Science and Technology, Tokyo, Japan (GRID:grid.26999.3d) (ISNI:0000 0001 2151 536X) 
6  Yonsei University College of Medicine, Graduate School of Medical Science, Brain Korea 21 Plus Project for Medical Sciences, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Yonsei University College of Medicine, Department and Research Institute of Rehabilitation Medicine, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Yonsei University, Graduate Program of Nano Science and Technology, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Yonsei University College of Medicine, Rehabilitation Institute of Neuromuscular Disease, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454) 
7  Seoul National University, Department of Electrical and Computer Engineering, Seoul, Republic of Korea (GRID:grid.31501.36) (ISNI:0000 0004 0470 5905); Seoul National University, Interdisciplinary Program in Artificial Intelligence, Seoul, Republic of Korea (GRID:grid.31501.36) (ISNI:0000 0004 0470 5905) 
8  Yonsei University College of Medicine, Department of Pharmacology, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Yonsei University College of Medicine, Graduate School of Medical Science, Brain Korea 21 Plus Project for Medical Sciences, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Yonsei University, Graduate Program of Nano Science and Technology, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Institute for Basic Science (IBS), Center for Nanomedicine, Seoul, Republic of Korea (GRID:grid.410720.0) (ISNI:0000 0004 1784 4496); Yonsei University, Graduate Program of Nano Biomedical Engineering (NanoBME), Advanced Science Institute, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454); Yonsei University College of Medicine, Severance Biomedical Science Institute, Seoul, Republic of Korea (GRID:grid.15444.30) (ISNI:0000 0004 0470 5454) 
Publication year
2021
Publication date
2021
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-021-25928-2
ProQuest document ID
2575662948
Copyright
© The Author(s) 2021. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.