| Chapter title |
The Nuclear Envelope
|
|---|---|
| Chapter number | 25 |
| Book title |
The Nuclear Envelope
|
| Published in |
Methods in molecular biology, January 2016
|
| DOI | 10.1007/978-1-4939-3530-7_25 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-3528-4, 978-1-4939-3530-7
|
| Authors |
Anton, Tobias, Leonhardt, Heinrich, Markaki, Yolanda, Tobias Anton, Heinrich Leonhardt, Yolanda Markaki |
| Editors |
Sue Shackleton, Philippe Collas, Eric C. Schirmer |
| Abstract |
The discovery that the RNA guided bacterial endonuclease Cas9 can be harnessed to target and manipulate user-defined genomic sequences has greatly influenced the field of genome engineering. Interestingly, a catalytically dead Cas9 (dCas9) can be employed as a targeted DNA-binding platform to alter gene expression. By fusing this dCas9 to eGFP, we and others could show that the CRISPR/Cas9 system can be further expanded to label and trace genomic loci in living cells. We demonstrated that by exchanging the sgRNA, dCas9-eGFP could be specifically directed to various heterochromatic sequences within the nucleus. Here, we provide a basic protocol for this versatile tool and describe how to verify new dCas9-eGFP targets. |
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 % |
|---|---|---|
| Netherlands | 1 | 3% |
| Unknown | 28 | 97% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 8 | 28% |
| Researcher | 5 | 17% |
| Student > Bachelor | 2 | 7% |
| Professor | 2 | 7% |
| Student > Doctoral Student | 1 | 3% |
| Other | 4 | 14% |
| Unknown | 7 | 24% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 12 | 41% |
| Agricultural and Biological Sciences | 8 | 28% |
| Immunology and Microbiology | 1 | 3% |
| Unknown | 8 | 28% |