Editing of DNA methylation using CRISPR/Cas9 and a ssDNA template in human cells.
Biochem Biophys Res Commun
; 581: 20-24, 2021 12 03.
Article
en En
| MEDLINE
| ID: mdl-34653674
ABSTRACT
Programmable DNA methylation is required for understanding of transcriptional regulation and elucidating gene functions. We previously reported that MMEJ-based promoter replacement enabled targeted DNA methylation in human cells. ssDNA-mediated knock-in has recently been reported to completely reduce random integrations. We speculated that by changing MMEJ-to ssDNA-based knock-in, targeted DNA methylation may be achieved through a hemimethylation-symmetric methylation pathway. We herein successfully developed a new system that enables the replacement of an unmethylated promoter with a methylated ssDNA promoter through ssDNA-based knock-in. A DNA methylation ratio of approximately 100% was achieved at the cancer-associated gene SP3 in HEK293 cells. The present results provide a promising framework for artificial epigenetic modifications.
Palabras clave
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
ADN de Cadena Simple
/
Metilación de ADN
/
Epigénesis Genética
/
Factor de Transcripción Sp3
/
Edición Génica
/
Proteínas de Neoplasias
Límite:
Humans
Idioma:
En
Revista:
Biochem Biophys Res Commun
Año:
2021
Tipo del documento:
Article