RNA biology in a test tube-an overview of in vitro systems/assays
In vitro systems have provided a wealth of information in the field of RNA biology, as they constitute a superior and sometimes the unique approach to address many important questions. Such cell-free methods can be sorted by the degree of complexity of the preparation of enzymatic and/or regulatory...
| Main Authors: | , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2013
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/97048 http://hdl.handle.net/10220/11748 |
| _version_ | 1826112038594674688 |
|---|---|
| author | Roca, Xavier Karginov, Fedor V. |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Roca, Xavier Karginov, Fedor V. |
| author_sort | Roca, Xavier |
| collection | NTU |
| description | In vitro systems have provided a wealth of information in the field of RNA biology, as they constitute a superior and sometimes the unique approach to address many important questions. Such cell-free methods can be sorted by the degree of complexity of the preparation of enzymatic and/or regulatory activity. Progress in the study of pre-mRNA processing has largely relied on traditional in vitro methods, as these reactions have been recapitulated in cell-free systems. The pre-mRNA capping, editing, and cleavage/polyadenylation reactions have even been reconstituted using purified components, and the enzymes responsible for catalysis have been characterized by such techniques. In vitro splicing using nuclear or cytoplasmic extracts has yielded clues on spliceosome assembly, kinetics, and mechanisms of splicing and has been essential to elucidate the function of splicing factors. Coupled systems have been important to functionally connect distinct processes, like transcription and splicing. Extract preparation has also been adapted to cells from a variety of tissues and species, revealing general versus species-specific mechanisms. Cell-free assays have also been applied to newly discovered pathways such as those involving small RNAs, including microRNAs (miRNAs), small interfering RNAs (siRNAs), and Piwi-interacting RNAs (piRNAs). The first two pathways have been well characterized largely by in vitro methods, which need to be developed for piRNAs. Finally, new techniques, such as single-molecule studies, are continuously being established, providing new and important insights into the field. Thus, in vitro approaches have been, are, and will continue being at the forefront of RNA research. |
| first_indexed | 2024-10-01T03:00:43Z |
| format | Journal Article |
| id | ntu-10356/97048 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:00:43Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | ntu-10356/970482020-03-07T12:18:18Z RNA biology in a test tube-an overview of in vitro systems/assays Roca, Xavier Karginov, Fedor V. School of Biological Sciences DRNTU::Science::Biological sciences In vitro systems have provided a wealth of information in the field of RNA biology, as they constitute a superior and sometimes the unique approach to address many important questions. Such cell-free methods can be sorted by the degree of complexity of the preparation of enzymatic and/or regulatory activity. Progress in the study of pre-mRNA processing has largely relied on traditional in vitro methods, as these reactions have been recapitulated in cell-free systems. The pre-mRNA capping, editing, and cleavage/polyadenylation reactions have even been reconstituted using purified components, and the enzymes responsible for catalysis have been characterized by such techniques. In vitro splicing using nuclear or cytoplasmic extracts has yielded clues on spliceosome assembly, kinetics, and mechanisms of splicing and has been essential to elucidate the function of splicing factors. Coupled systems have been important to functionally connect distinct processes, like transcription and splicing. Extract preparation has also been adapted to cells from a variety of tissues and species, revealing general versus species-specific mechanisms. Cell-free assays have also been applied to newly discovered pathways such as those involving small RNAs, including microRNAs (miRNAs), small interfering RNAs (siRNAs), and Piwi-interacting RNAs (piRNAs). The first two pathways have been well characterized largely by in vitro methods, which need to be developed for piRNAs. Finally, new techniques, such as single-molecule studies, are continuously being established, providing new and important insights into the field. Thus, in vitro approaches have been, are, and will continue being at the forefront of RNA research. 2013-07-17T06:46:21Z 2019-12-06T19:38:13Z 2013-07-17T06:46:21Z 2019-12-06T19:38:13Z 2012-07 2012 Journal Article Roca, X., & Karginov, F. V. (2012). RNA biology in a test tube-an overview of in vitro systems/assays. Wiley Interdisciplinary Reviews: RNA, 3(4), 509-527. 1757-7012 https://hdl.handle.net/10356/97048 http://hdl.handle.net/10220/11748 10.1002/wrna.1115 en Wiley interdisciplinary reviews : RNA © 2012 John Wiley & Sons, Ltd. |
| spellingShingle | DRNTU::Science::Biological sciences Roca, Xavier Karginov, Fedor V. RNA biology in a test tube-an overview of in vitro systems/assays |
| title | RNA biology in a test tube-an overview of in vitro systems/assays |
| title_full | RNA biology in a test tube-an overview of in vitro systems/assays |
| title_fullStr | RNA biology in a test tube-an overview of in vitro systems/assays |
| title_full_unstemmed | RNA biology in a test tube-an overview of in vitro systems/assays |
| title_short | RNA biology in a test tube-an overview of in vitro systems/assays |
| title_sort | rna biology in a test tube an overview of in vitro systems assays |
| topic | DRNTU::Science::Biological sciences |
| url | https://hdl.handle.net/10356/97048 http://hdl.handle.net/10220/11748 |
| work_keys_str_mv | AT rocaxavier rnabiologyinatesttubeanoverviewofinvitrosystemsassays AT karginovfedorv rnabiologyinatesttubeanoverviewofinvitrosystemsassays |