| Chapter title |
In vivo screening of ligand-dependent hammerhead ribozymes.
|
|---|---|
| Chapter number | 28 |
| Book title |
Ribozymes
|
| Published in |
Methods in molecular biology, January 2012
|
| DOI | 10.1007/978-1-61779-545-9_28 |
| Pubmed ID | |
| Book ISBNs |
978-1-61779-544-2, 978-1-61779-545-9
|
| Authors |
Athanasios Saragliadis, Benedikt Klauser, Jörg S. Hartig, Saragliadis, Athanasios, Klauser, Benedikt, Hartig, Jörg S. |
| Abstract |
The development of artificial switches of gene expression is of high importance for future applications in biotechnology and synthetic biology. We have developed a powerful RNA-based system which allows for the ligand-dependent and reprogrammable control of gene expression in Escherichia coli. Our system makes use of the hammerhead ribozyme (HHR) which acts as molecular scaffold for the sequestration of the ribosome binding site (RBS), mimicking expression platforms in naturally occurring riboswitches. Aptamer domains can be attached to the ribozyme as exchangeable ligand-sensing modules. Addition of ligands to the bacterial growth medium changes the activity of the ligand-dependent self-cleaving ribozyme which in turn switches gene expression. In this chapter, we describe the in vivo screening procedure allowing for reprogramming the ligand-specificity of our system. |
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 % |
|---|---|---|
| United States | 1 | 6% |
| China | 1 | 6% |
| Unknown | 15 | 88% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 4 | 24% |
| Student > Master | 3 | 18% |
| Researcher | 3 | 18% |
| Student > Bachelor | 1 | 6% |
| Other | 1 | 6% |
| Other | 1 | 6% |
| Unknown | 4 | 24% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 7 | 41% |
| Biochemistry, Genetics and Molecular Biology | 4 | 24% |
| Chemistry | 1 | 6% |
| Medicine and Dentistry | 1 | 6% |
| Unknown | 4 | 24% |