| Chapter title |
In Vivo Production of Small Recombinant RNAs Embedded in a 5S rRNA-Derived Protective Scaffold
|
|---|---|
| Chapter number | 5 |
| Book title |
RNA Scaffolds
|
| Published in |
Methods in molecular biology, January 2015
|
| DOI | 10.1007/978-1-4939-2730-2_5 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-2729-6, 978-1-4939-2730-2
|
| Authors |
Victor G. Stepanov, George E. Fox, Stepanov, Victor G., Fox, George E. |
| Abstract |
Preparative synthesis of RNA is a challenging task that is usually accomplished using either chemical or enzymatic polymerization of ribonucleotides in vitro. Herein, we describe an alternative approach in which RNAs of interest are expressed as a fusion with a 5S rRNA-derived scaffold. The scaffold provides protection against cellular ribonucleases resulting in cellular accumulations comparable to those of regular ribosomal RNAs. After isolation of the chimeric RNA from the cells, the scaffold can be removed if necessary by deoxyribozyme-catalyzed cleavage followed by preparative electrophoretic separation of the cleavage reaction products. The protocol is designed for sustained production of high quality RNA on the milligram scale. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| United States | 1 | 20% |
| Unknown | 4 | 80% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 2 | 40% |
| Researcher | 2 | 40% |
| Student > Bachelor | 1 | 20% |
| Readers by discipline | Count | As % |
|---|---|---|
| Agricultural and Biological Sciences | 3 | 60% |
| Biochemistry, Genetics and Molecular Biology | 2 | 40% |