| Chapter title |
Conditional Toxin Splicing Using a Split Intein System
|
|---|---|
| Chapter number | 13 |
| Book title |
Split Inteins
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6451-2_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6449-9, 978-1-4939-6451-2
|
| Authors |
Spencer C. Alford, Connor O’Sullivan, Perry L. Howard |
| Abstract |
Protein toxin splicing mediated by split inteins can be used as a strategy for conditional cell ablation. The approach requires artificial fragmentation of a potent protein toxin and tethering each toxin fragment to a split intein fragment. The toxin-intein fragments are, in turn, fused to dimerization domains, such that addition of a dimerizing agent reconstitutes the split intein. These chimeric toxin-intein fusions remain nontoxic until the dimerizer is added, resulting in activation of intein splicing and ligation of toxin fragments to form an active toxin. Considerations for the engineering and implementation of conditional toxin splicing (CTS) systems include: choice of toxin split site, split site (extein) chemistry, and temperature sensitivity. The following method outlines design criteria and implementation notes for CTS using a previously engineered system for splicing a toxin called sarcin, as well as for developing alternative CTS systems. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 5 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Librarian | 1 | 20% |
| Professor > Associate Professor | 1 | 20% |
| Researcher | 1 | 20% |
| Student > Doctoral Student | 1 | 20% |
| Unknown | 1 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 2 | 40% |
| Biochemistry, Genetics and Molecular Biology | 1 | 20% |
| Medicine and Dentistry | 1 | 20% |
| Unknown | 1 | 20% |