Advances in base editing with an emphasis on an AAV-based strategy

Jiajie Kuang; Qinghua Lyu; Jiao Wang; Yubo Cui; Jun Zhao

doi:10.1016/j.ymeth.2021.03.015

Advances in base editing with an emphasis on an AAV-based strategy

Methods. 2021 Oct:194:56-64. doi: 10.1016/j.ymeth.2021.03.015. Epub 2021 Mar 25.

Authors

Jiajie Kuang¹, Qinghua Lyu², Jiao Wang³, Yubo Cui⁴, Jun Zhao⁵

Affiliations

¹ Shenzhen Eye Institute, Shenzhen Eye Hospital, Jinan University, Shenzhen 518000, China; Department of Ophthalmology, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China.
² School of Ophthalmology & Optometry, Shenzhen Eye Hospital, Shenzhen University, Shenzhen 518000, China; Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, China.
³ School of Ophthalmology & Optometry, Shenzhen Eye Hospital, Shenzhen University, Shenzhen 518000, China.
⁴ Department of Ophthalmology, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China.
⁵ Shenzhen Eye Institute, Shenzhen Eye Hospital, Jinan University, Shenzhen 518000, China; Department of Ophthalmology, Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China. Electronic address: doctorzhaojun@163.com.

PMID: 33774157
DOI: 10.1016/j.ymeth.2021.03.015

Abstract

The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based base editors have been developed for precisely installing point mutations in genomes with high efficiency. Two editing systems of cytosine base editors (CBEs) and adenine base editors (ABEs) have been developed for conversion of C.G-to-T.A and A.T-to-G.C, respectively, showing the prominence in genomic DNA correction and mutation. Here, we summarize recent optimized approaches in improving base editors, including the evolution of Cas proteins, the choice of deamination enzymes, modification on linker length, base-editor expression, and addition of functional domains. Specifically, in this paper we highlight a strategy of split-intein mediated base-editor reconstitution for its adeno-associated virus (AAV) delivery. The purpose of this article is to offer readers with a better understanding of AAV-mediated base editors, and facilitate them to use this tool in in vivo experiments and potential clinical applications.

Keywords: AAV delivery; Base editors; CRISPR; Editing efficiency.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics
Cytosine
DNA
Dependovirus / genetics
Gene Editing*

Substances

Cytosine
DNA