Evaluating and Enhancing Target Specificity of Gene-Editing Nucleases and Deaminases.
Annu Rev Biochem
; 88: 191-220, 2019 06 20.
Article
em 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.
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Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Engenharia de Proteínas
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RNA Guia de Cinetoplastídeos
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Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
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Sistemas CRISPR-Cas
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Edição de Genes
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Proteína 9 Associada à CRISPR
Limite:
Humans
Idioma:
En
Revista:
Annu Rev Biochem
Ano de publicação:
2019
Tipo de documento:
Article