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Updated: Aug 14, 2025

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A CRISPR/Cas9-based method for targeted DNA methylation enables cancer initiation in B lymphocytes

Shota Katayama1, Koichi Shiraishi1,2, Naoki Gorai1,2

  • 1IMRA Japan Co., Ltd. Sapporo Japan.

Advanced Genetics (Hoboken, N.J.)
|January 9, 2023
PubMed

Insights

Researchers developed a novel CRISPR-Cas9 system for targeted DNA methylation, successfully inducing cancer-associated gene hypermethylation and B-cell lymphoma in human cells.

Area of Science:

  • Epigenetics and molecular biology
  • Cancer research
  • Gene regulation

Background:

  • Targeted DNA methylation is crucial for understanding gene transcription and epigenetic diseases.
  • CRISPR-Cas9 technology holds promise for DNA methylation but faces efficiency challenges in inducing epigenetic alterations.

Purpose of the Study:

  • To develop an efficient system for targeted DNA methylation using CRISPR-Cas9.
  • To investigate the induction of epigenetic diseases through engineered DNA methylation.

Main Methods:

  • Developed a novel microhomology-mediated end joining (MHEJ)-based knock-in system.
  • Replaced unmethylated promoters with methylated ones.
  • Utilized CRISPR-Cas9 for targeted genomic modifications.

Main Results:

  • Achieved approximately 100% DNA methylation at the SP3 gene promoter in HEK293 cells.
  • Engineered SP3 promoter hypermethylation resulted in transcriptional suppression in human B lymphocytes.
  • Induced B-cell lymphoma through targeted epimutations.

Conclusions:

  • The developed system enables efficient targeted DNA methylation.
  • This approach offers a promising framework for studying cancer initiation via epimutations.
  • The system can be used to model and potentially treat epigenetic diseases.
Keywords:
CRISPR/Cas9DNA methylationMMEJcancer initiationcancer risk evaluationgene expression

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