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CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

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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

CRISPR/Cas9-Based Engineering of the Epigenome.

Julian Pulecio¹, Nipun Verma², Eva Mejía-Ramírez³

¹Center of Regenerative Medicine in Barcelona (CMRB), Hospital Duran i Reynals, 3rd floor, Avenue Gran Via 199-203, Hospitalet de Llobregat, 08908 Barcelona, Spain; Center for Networked Biomedical Research on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), 28029 Madrid, Spain; Developmental Biology Program, Sloan Kettering Institute, 1275 York Avenue, New York, NY 10065, USA.

|October 7, 2017

Summary

CRISPR/Cas9 technology revolutionizes epigenome editing, allowing scientists to directly link chromatin modifications to gene expression and cell behavior. This advancement aids stem cell research and therapeutic engineering.

Keywords:

CRISPR-dCas9 DNA methylation chromatin architecture histone modifications targeted epigenome engineering targeted gene edition transcriptional regulation

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Area of Science:

Epigenetics and Molecular Biology
Stem Cell Biology
Gene Editing Technologies

Background:

Establishing causal links between chromatin features and gene expression/cell behavior is challenging.
Epigenome editing tools are emerging, enabling locus-specific chromatin modification analysis.
Understanding these relationships is crucial for stem cell biology and therapeutic applications.

Purpose of the Study:

To review the potential of CRISPR/Cas9 technology for epigenome investigation and manipulation.
To discuss the application of epigenome engineering in stem cell biology and therapeutic development.
To highlight technical considerations for improving CRISPR/Cas9-based epigenome engineering tools.

Main Methods:

Review of CRISPR/Cas9-based epigenome editing strategies.
Discussion of methods for assigning functional consequences to chromatin modifications.
Analysis of technical aspects for standardization and advancement of epigenome engineering tools.

Main Results:

CRISPR/Cas9 offers a powerful platform for targeted epigenome manipulation.
This technology enables direct assessment of locus-specific chromatin modification impacts.
Advancements facilitate deeper understanding of stem cell regulation and engineering potential.

Conclusions:

CRISPR/Cas9 technology provides an unprecedented opportunity to study and engineer the epigenome.
This approach is vital for advancing stem cell biology and developing cell-based therapies.
Further technical refinement is needed to optimize CRISPR/Cas9 tools for epigenome engineering.