Understanding the diversity of genetic outcomes from CRISPR-Cas generated homology-directed repair.
Commun Biol
; 2: 458, 2019.
Article
en En
| MEDLINE
| ID: mdl-31840103
ABSTRACT
As CRISPR-Cas systems advance toward clinical application, it is essential to identify all the outcomes of gene-editing activity in human cells. Reports highlighting the remarkable success of homology-directed repair (HDR) in the treatment of inherited diseases may inadvertently underreport the collateral activity of this remarkable technology. We are utilizing an in vitro gene-editing system in which a CRISPR-Cas complex provides the double-stranded cleavage and a mammalian cell-free extract provides the enzymatic activity to promote non-homologous end joining, micro-homology mediated end joining, and homology-directed repair. Here, we detail the broad spectrum of gene-editing reaction outcomes utilizing Cas9 and Cas12a in combination with single-stranded donor templates of the sense and nonsense polarity. This system offers the opportunity to see the range of outcomes of gene-editing reactions in an unbiased fashion, detailing the distribution of DNA repair outcomes as a function of a set of genetic tools.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Variación Genética
/
Reparación del ADN
/
Sistemas CRISPR-Cas
/
Edición Génica
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Commun Biol
Año:
2019
Tipo del documento:
Article