Application of CRISPR-Cas9 in Epigenome Editing    

Epigenetic regulatory mechanisms play a central role in almost all cellular phenomena by converting internal and environmental signals into gene transcription. Breakthroughs in CRISPR/Cas-based epigenetic editing technologies have enabled researchers to target specific sites for epigenetic modifications, thereby manipulating the structure of endogenous DNA and histones and changing the architecture of native chromatin. Inactive CRISPR/Cas systems (dCas) without nuclease activity have been repurposed as synthetic DNA binding platforms. These platforms have been used to reorganize chromatin structure and recruit effectors that alter the epigenetic group and gene expression at specific sites. Since genomic DNA is relatively simple to target by changing the in situ spacer sequence in the guide RNA (gRNA), dCas-based effectors have revolutionized our ability to edit the epigenetic group and greatly increased our understanding of epigenetic regulation.

Technical Application Direction

Precise Regulation of Gene Expression

The application of CRISPR/Cas9 technology in epigenetics can achieve precise regulation of genes, including activation and inhibition of the expression level of specific genes. This helps to reveal gene function and regulatory mechanisms and further understand the molecular mechanisms of disease occurrence and development.

Establish Disease Models

By using CRISPR/Cas9 technology to construct epigenetic-related disease models in cell and animal models, the process of disease occurrence can be simulated more accurately, and new ideas and methods can be provided for the study and treatment of diseases.

Find Therapeutic Targets

Gene editing and epigenetic regulation using CRISPR/Cas9 technology can help identify possible therapeutic targets and create tailored medicines, offering fresh approaches to illness treatment.

Based on the individual's genomic and epigenetic characteristics, using CRISPR/Cas9 technology for precise gene editing and regulation can achieve the goal of personalized medicine and tailor the most appropriate treatment plan for each patient.

Figure 1. Nuclease-deficient CRISPR/Cas systems serve as platforms for epigenome control across various scales, enabling programmable manipulation of genomic organization, chromatin looping, and biochemical modifications. (Goell JH, et al., 2021)

Technical Principle

The core of CRISPR/Cas-based epigenome editing technology is to use the dCas protein in the CRISPR/Cas system for site-specific binding of DNA, and to locate the dCas protein to a specific genomic region through guide RNA (gRNA). Unlike conventional genome editing, this method does not involve direct modification of DNA sequences but rather regulates the expression of target genes by changing chromatin structure or introducing specific epigenetic modifications.

Technical Details

Choice of dCas Protein

In the CRISPR/Cas system, the dCas protein is an inactivated Cas9 protein that does not have nuclease activity but can still bind to the gRNA and locate to the target DNA sequence. This makes the dCas protein an ideal tool for epigenetic editing because it can achieve site-specific directional regulation of genomic regions without inducing DNA double-stranded cleavage.

gRNA Design

gRNA is a key component that binds to the dCas protein and directs it to the target genomic region. The design of the gRNA needs to take into account its compatibility with the target DNA sequence and potential off-target effects. By rationally designing the gRNA sequence, efficient epigenetic editing can be achieved and non-specific modifications can be minimized.

Choice of Epigenetic Effector

In CRISPR/Cas-based epigenome editing, the effector used determines how chromatin structure and gene expression are regulated. Commonly used effectors include DNA methyltransferases (such as DNMTs), histone acetyltransferases (such as p300/CBP), histone deacetylases (such as HDACs), etc. Selecting the right effector can achieve precise modification of the target genomic region, thereby regulating gene expression levels.

Complex Multiple Effectors

In order to achieve simultaneous regulation of multiple genes in a single cell, researchers are exploring methods to combine multiple effectors into the same CRISPR/Cas complex. Such a complex can simultaneously regulate multiple epigenetic modification pathways, thereby achieving precise regulation of complex gene networks.

Spatiotemporal Regulation Strategies

Achieving spatiotemporal regulation of epigenetic editing systems is essential for their safety and effectiveness in clinical applications. By introducing tissue-specific and conditional promoters, using anti-CRISPR proteins, small molecule inducers, and protease-activated Cas variants, precise spatiotemporal control of the editing process can be achieved, thereby minimizing off-target effects and ensuring editing efficiency.

Creative Biogene's CRISPR services enable you to harness the potential of precise epigenetic editing! Our cutting-edge CRISPR/Cas9 technology allows for precise regulation of gene expression, providing unrivaled control over cellular function. Our expertise in CRISPR-based epigenome editing enables innovative research and therapeutic development, from disease modeling to therapeutic target discovery and personalized medicine. Collaborate with us to transform your research and expedite your road to scientific discovery.

Related Services and Products

Custom Cell Lines Services
sgRNA Bioinformatics Service
SgRNA Design and Confirmation
Base Editing by CRISPR