Chapter title |
Editing of DNA Methylation Using dCas9-Peptide Repeat and scFv-TET1 Catalytic Domain Fusions
|
---|---|
Chapter number | 23 |
Book title |
Epigenome Editing
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7774-1_23 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7773-4, 978-1-4939-7774-1
|
Authors |
Sumiyo Morita, Takuro Horii, Izuho Hatada, Morita, Sumiyo, Horii, Takuro, Hatada, Izuho |
Abstract |
DNA methylation, one of the most studied epigenetic modifications, regulates many biological processes. Dysregulation of DNA methylation is implicated in the etiology of several diseases, such as cancer and imprinting diseases. Accordingly, technologies designed to manipulate DNA methylation at specific loci are very important, and many epigenome editing technologies have been developed, based on zinc finger proteins, TALEs, and CRISPR/dCas9 targeting. We describe a protocol to induce and assess DNA demethylation on a target gene. It is based on a modification of the dCas9-SunTag system for efficient, targeted demethylation at specific DNA loci. The original SunTag system consists of ten copies of the GCN4 peptide separated by 5-amino-acid linkers. To achieve efficient recruitment of an anti-GCN4 scFv fused to the ten-eleven (TET) 1 hydroxylase, an enzyme that demethylates DNA, we changed the linker length to 22 amino acids. |
X Demographics
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 1 | 100% |
Demographic breakdown
Type | Count | As % |
---|---|---|
Members of the public | 1 | 100% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
Unknown | 25 | 100% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Student > Bachelor | 5 | 20% |
Researcher | 4 | 16% |
Other | 2 | 8% |
Student > Ph. D. Student | 2 | 8% |
Student > Doctoral Student | 1 | 4% |
Other | 4 | 16% |
Unknown | 7 | 28% |
Readers by discipline | Count | As % |
---|---|---|
Biochemistry, Genetics and Molecular Biology | 11 | 44% |
Agricultural and Biological Sciences | 2 | 8% |
Linguistics | 1 | 4% |
Nursing and Health Professions | 1 | 4% |
Medicine and Dentistry | 1 | 4% |
Other | 0 | 0% |
Unknown | 9 | 36% |