RNA biology in a test tubeuan overview of in vitro systems/assays

Roca, X., Karginov, F. V. (July 2012) RNA biology in a test tubeuan overview of in vitro systems/assays. Wiley Interdisciplinary Reviews-Rna, 3 (4). pp. 509-527. ISSN 1757-7004

URL: http://www.ncbi.nlm.nih.gov/pubmed/22447682
DOI: 10.1002/wrna.1115


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. WIREs RNA 2012, 3:509527. doi: 10.1002/wrna.1115 For further resources related to this article, please visit the .

Item Type: Paper
Uncontrolled Keywords: pre-messenger-rna 5' splice-site small nuclear ribonucleoproteins polyadenylation factor cpsf-73 cap-binding complex polymerase-ii saccharomyces-cerevisiae poly(a) polymerase decapping enzyme capping enzyme
Subjects: bioinformatics
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics
Investigative techniques and equipment
CSHL Authors:
Communities: CSHL labs > Hannon lab
CSHL Post Doctoral Fellows
Depositing User: Matt Covey
Date: July 2012
Date Deposited: 18 Jan 2013 20:15
Last Modified: 02 May 2013 19:35
Related URLs:
URI: https://repository.cshl.edu/id/eprint/27031

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