CRISPR/Cas9-mediated epigenetic editing tool: An optimized strategy for targeting de novo DNA methylation with stable status via homology directed repair pathway

Upregulation of DNA methyltransferases (DNMTs) was observed in edited HeLa cells.

• Targeted methylation of the MGMT gene was achieved using CRISPR/Cas9-mediated repair techniques.
• Transient methylation levels in edited cells were identified as a challenge for stable gene regulation.
• An optimized approach using the DNA ligase IV inhibitor SCR7 improved insertion efficiency of methylated repair templates.
• The combination of CRISPR/Cas9 components, SCR7 treatment, and long methylated templates may enhance epigenomic editing efficiency.
• Site-specific methylation facilitated by these methods could maintain gene silencing in both laboratory and living systems.

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