CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation
CRISPR/Cas9-mediated cleavage of DNA, which depends on the endonuclease activity of Cas9, has been widely used for gene editing due to its excellent programmability and specificity. However, the changes to the DNA sequence that are mediated by CRISPR/Cas9 affect the structures and stability of the g...
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| Language: | English |
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MDPI AG
2023-10-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/24/19/14865 |
| _version_ | 1797575761516822528 |
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| author | Ruijie Cai Runyu Lv Xin’e Shi Gongshe Yang Jianjun Jin |
| author_facet | Ruijie Cai Runyu Lv Xin’e Shi Gongshe Yang Jianjun Jin |
| author_sort | Ruijie Cai |
| collection | DOAJ |
| description | CRISPR/Cas9-mediated cleavage of DNA, which depends on the endonuclease activity of Cas9, has been widely used for gene editing due to its excellent programmability and specificity. However, the changes to the DNA sequence that are mediated by CRISPR/Cas9 affect the structures and stability of the genome, which may affect the accuracy of results. Mutations in the RuvC and HNH regions of the Cas9 protein lead to the inactivation of Cas9 into dCas9 with no endonuclease activity. Despite the loss of endonuclease activity, dCas9 can still bind the DNA strand using guide RNA. Recently, proteins with active/inhibitory effects have been linked to the end of the dCas9 protein to form fusion proteins with transcriptional active/inhibitory effects, named CRISPRa and CRISPRi, respectively. These CRISPR tools mediate the transcription activity of protein-coding and non-coding genes by regulating the chromosomal modification states of target gene promoters, enhancers, and other functional elements. Here, we highlight the epigenetic mechanisms and applications of the common CRISPR/dCas9 tools, by which we hope to provide a reference for future related gene regulation, gene function, high-throughput target gene screening, and disease treatment. |
| first_indexed | 2024-03-10T21:42:54Z |
| format | Article |
| id | doaj.art-97bf9d7c7cc84734850369c055132347 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T21:42:54Z |
| publishDate | 2023-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-97bf9d7c7cc84734850369c0551323472023-11-19T14:31:48ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672023-10-0124191486510.3390/ijms241914865CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional RegulationRuijie Cai0Runyu Lv1Xin’e Shi2Gongshe Yang3Jianjun Jin4Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, ChinaLaboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, ChinaLaboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, ChinaLaboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, ChinaLaboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, ChinaCRISPR/Cas9-mediated cleavage of DNA, which depends on the endonuclease activity of Cas9, has been widely used for gene editing due to its excellent programmability and specificity. However, the changes to the DNA sequence that are mediated by CRISPR/Cas9 affect the structures and stability of the genome, which may affect the accuracy of results. Mutations in the RuvC and HNH regions of the Cas9 protein lead to the inactivation of Cas9 into dCas9 with no endonuclease activity. Despite the loss of endonuclease activity, dCas9 can still bind the DNA strand using guide RNA. Recently, proteins with active/inhibitory effects have been linked to the end of the dCas9 protein to form fusion proteins with transcriptional active/inhibitory effects, named CRISPRa and CRISPRi, respectively. These CRISPR tools mediate the transcription activity of protein-coding and non-coding genes by regulating the chromosomal modification states of target gene promoters, enhancers, and other functional elements. Here, we highlight the epigenetic mechanisms and applications of the common CRISPR/dCas9 tools, by which we hope to provide a reference for future related gene regulation, gene function, high-throughput target gene screening, and disease treatment.https://www.mdpi.com/1422-0067/24/19/14865CRISPR activationCRISPR inhibitionhistone modificationDNA methylationtranscription regulation |
| spellingShingle | Ruijie Cai Runyu Lv Xin’e Shi Gongshe Yang Jianjun Jin CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation International Journal of Molecular Sciences CRISPR activation CRISPR inhibition histone modification DNA methylation transcription regulation |
| title | CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation |
| title_full | CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation |
| title_fullStr | CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation |
| title_full_unstemmed | CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation |
| title_short | CRISPR/dCas9 Tools: Epigenetic Mechanism and Application in Gene Transcriptional Regulation |
| title_sort | crispr dcas9 tools epigenetic mechanism and application in gene transcriptional regulation |
| topic | CRISPR activation CRISPR inhibition histone modification DNA methylation transcription regulation |
| url | https://www.mdpi.com/1422-0067/24/19/14865 |
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