Related Experiment Video
Updated: Jul 1, 2025
07:49
CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
Published on: May 30, 2025
917
Targeted Modification of Epigenetic Marks Using CRISPR/dCas9-SunTag-Based Modular Epigenetic Toolkit
Min Kyung Song1,2, Yoon-Seong Kim3
1RWJMS Institute for Neurological Therapeutics, Rutgers-Robert Wood Johnson Medical School, Piscataway, NJ, USA.
Methods in Molecular Biology (Clifton, N.J.)
|March 1, 2024
Summary
Researchers developed a CRISPR/dCas9-SunTag toolkit for precise gene modulation. This system allows detailed study of epigenetic regulation, advancing understanding of gene expression control.
Area of Science:
- Epigenetics
- Molecular Biology
- Gene Regulation
Background:
- The epigenome comprises DNA and histone modifications influencing gene expression.
- CRISPR/dCas9 systems offer targeted gene modulation via effector domains.
- CRISPR/dCas9-SunTag enhances gene activation/repression by recruiting multiple proteins.
Purpose of the Study:
- To develop a modular epigenetic toolkit for gene-specific epigenetic architecture modulation.
- To establish a stable cell line for studying H3K4Me3-mediated promoter regulation at high resolution.
Main Methods:
- Designing sgRNAs and subcloning dCas9-5XGCN4 into pLvx-DsRed.
- Utilizing CRISPR/dCas9-SunTag-JARID1A system in SH-SY5Y cells.
- Validating epigenetic mark changes using ChIP and gene expression using RT-qPCR.
Main Results:
- Successful generation of a stable cell line for epigenetic studies.
- Demonstrated ability to modulate gene-specific epigenetic architecture.
- High-resolution analysis of H3K4Me3-mediated promoter regulation.
Conclusions:
- The developed CRISPR/dCas9-SunTag toolkit enables precise gene-specific epigenetic modulation.
- This toolkit is valuable for investigating the link between epigenetic modifications and gene expression.
- Facilitates high-resolution studies of epigenetic regulation in cellular systems.
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