Evaluating and Enhancing Target Specificity of Gene-Editing Nucleases and Deaminases.
Annu Rev Biochem
; 88: 191-220, 2019 06 20.
Article
in En
| MEDLINE
| ID: mdl-30883196
ABSTRACT
Programmable nucleases and deaminases, which include zinc-finger nucleases, transcription activator-like effector nucleases, CRISPR RNA-guided nucleases, and RNA-guided base editors, are now widely employed for the targeted modification of genomes in cells and organisms. These gene-editing tools hold tremendous promise for therapeutic applications. Importantly, these nucleases and deaminases may display off-target activity through the recognition of near-cognate DNA sequences to their target sites, resulting in collateral damage to the genome in the form of local mutagenesis or genomic rearrangements. For therapeutic genome-editing applications with these classes of programmable enzymes, it is essential to measure and limit genome-wide off-target activity. Herein, we discuss the key determinants of off-target activity for these systems. We describe various cell-based and cell-free methods for identifying genome-wide off-target sites and diverse strategies that have been developed for reducing the off-target activity of programmable gene-editing enzymes.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Protein Engineering
/
RNA, Guide, Kinetoplastida
/
Clustered Regularly Interspaced Short Palindromic Repeats
/
CRISPR-Cas Systems
/
Gene Editing
/
CRISPR-Associated Protein 9
Limits:
Humans
Language:
En
Journal:
Annu Rev Biochem
Year:
2019
Type:
Article