CRISPRoff/CRISPRon Service

Gene editing is a new technology for site-specific modification of the genome. Gene editing technology can cut the target DNA segment at a specific site, re-write, remove, or add the gene segment at a specific location. Although gene editing technology has shown great potential in gene research, disease prevention and genetic improvement, editing involves the cutting of DNA strands, which will cause permanent changes in the genetic material of cells. Recently, CRISPRoff/CRISPRon, a emerging programmable epigenomeediting technology, can solve this problem, and control gene expression with high specificity without changing the DNA sequence.

Figure 1. Working Mechanism of CRISPR off and on technology. (Nuez et al. 2021)

CRISPRoff is a programmable epigenetic memory writer that mainly promotes DNA methylation near the target site to achieve sustained heritable gene silencing. In the CRISPRoff system, sgRNA can guide Cas9 to the target site, which is a single dead Cas9 (dCas9) fusion protein that establishes DNA methylation and repressive histone modifications. At the same time, the CRISPRon system can reverse the editing of CRISPRoff. The principle of CRISPRon is similar to that of CRISPRoff. CRISPRon also utilizes the positioning ability of the combination of sgRNA and dCas9, except that DNA demethylase is fused to dCas9.

Advantages of the CRISPRoff/CRISPRon System.

• Flexibility and Stability. Expression of the protein encoded by the target gene was detected to decrease significantly trough CRISPRoff system. Besides, mRNA transcribed by the target gene was reduced, and the DNA methylation of its CpG island increased.
• Specificity and Wide Applicability. CRISPRoff does not affect the epigenetic sequence adjacent to the target site status. The editing effect is resistant to cell division and differentiation.
• Recoverability. After CRISPRon treatment, 74% of cells recovered the expression of the target gene, and the methylation of the CpG island of the target gene basically disappeared.

Figure 2. CRISPRoff-mediated silencing of genes without promoter CpG island annotations. (Nuez et al. 2021)

Lifeasible is a world-class phytotechnology company with extensive experience in plant gene editing. Based on mature and efficient CRISPR system, we are committed to developing epigenetic editor to better serve customers. Our epigenetic editor services are proudly featured with high quality and efficiency. We hope that our technology and services can help you achieve new breakthroughs. For more information, please contact Lifeasible.

CRISPRoff /CRISPRon service workflow