| Chapter title |
Purification and In Vitro Characterization of Zinc Finger Recombinases
|
|---|---|
| Chapter number | 15 |
| Book title |
Site-Specific Recombinases
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-7169-5_15 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7167-1, 978-1-4939-7169-5
|
| Authors |
Femi J. Olorunniji, Susan J. Rosser, W. Marshall Stark, Olorunniji, Femi J., Rosser, Susan J., Marshall Stark, W. |
| Abstract |
Zinc finger recombinases (ZFRs) are designer site-specific recombinases that have been adapted for a variety of genome editing purposes. Due to their modular nature, ZFRs can be customized for targeted sequence recognition and recombination. There has been substantial research on the in vivo properties and applications of ZFRs; however, in order to fully understand and customize them, it will be necessary to study their properties in vitro. Experiments in vitro can allow us to optimize catalytic activities, improve target specificity, measure and minimize off-target activity, and characterize key steps in the recombination pathway that might be modified to improve performance. Here, we present a straightforward set of protocols for the expression and purification of ZFRs, an assay system for catalytic proficiency in vitro and bandshift assays for detection of sequence-specific DNA interactions. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 6 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 3 | 50% |
| Student > Ph. D. Student | 1 | 17% |
| Student > Bachelor | 1 | 17% |
| Student > Postgraduate | 1 | 17% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 4 | 67% |
| Biochemistry, Genetics and Molecular Biology | 2 | 33% |