A CRISPR/Cas9-based method for targeted DNA methylation enables cancer initiation in B lymphocytes.
Shota KatayamaKoichi ShiraishiNaoki GoraiMasao AndouPublished in: Advanced genetics (Hoboken, N.J.) (2021)
Targeted DNA methylation is important for understanding transcriptional modulation and epigenetic diseases. Although CRISPR-Cas9 has potential for this purpose, it has not yet been successfully used to efficiently introduce DNA methylation and induce epigenetic diseases. We herein developed a new system that enables the replacement of an unmethylated promoter with a methylated promoter through microhomology-mediated end joining-based knock-in. We successfully introduced an approximately 100% DNA methylation ratio at the cancer-associated gene SP3 in HEK293 cells. Moreover, engineered SP3 promoter hypermethylation led to transcriptional suppression in human B lymphocytes and induced B-cell lymphoma. Our system provides a promising framework for targeted DNA methylation and cancer initiation through epimutations.
Keyphrases
- dna methylation
- genome wide
- gene expression
- crispr cas
- genome editing
- copy number
- papillary thyroid
- cancer therapy
- induced apoptosis
- peripheral blood
- endothelial cells
- transcription factor
- squamous cell
- squamous cell carcinoma
- signaling pathway
- drug delivery
- cell cycle arrest
- cell death
- diffuse large b cell lymphoma
- young adults
- risk assessment
- drug induced
- dna damage
- genome wide identification