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CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

Published on: May 30, 2025

Destabilization of the human epigenome: consequences of foreign DNA insertions

Stefanie Weber¹, Andrea Hofmann², Stefan Herms^2,3

¹Institute of Clinical & Molecular Virology, University of Erlangen-Nürnberg Medical School, D-91054 Erlangen, Germany.

|June 20, 2015

Abstract

Aim:

We previously reported changes of DNA methylation and transcription patterns in mammalian cells that carry integrated foreign DNA. Experiments were now designed to assess the epigenetic consequences of inserting a 5.6 kbp plasmid into the human genome.

Methods:

Differential transcription and CpG methylation patterns were compared between transgenomic and nontransgenomic cell clones by using gene chip microarray systems.

Results:

In 4.7% of the 28.869 gene segments analyzed, transcriptional activities were up- or downregulated in the transgenomic cell clones. Genome-wide profiling revealed differential methylation in 3791 of > 480,000 CpG's examined in transgenomic versus nontransgenomic clones.

Conclusion:

The data document genome-wide effects of foreign DNA insertions on the epigenetic stability of human cells. Many fields in experimental biology and medicine employ transgenomic or otherwise genome-manipulated cells or organisms without considering the epigenetic consequences for the recipient genomes.

Keywords:

DNA methylation destabilization differential gene expression differential methylation epigenetics foreign DNA transgenomes

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