Overview of Methods for Large-Scale RNA Synthesis
In recent years, it has become clear that RNA molecules are involved in almost all vital cellular processes and pathogenesis of human disorders. The functional diversity of RNA comes from its structural richness. Although composed of only four nucleotides, RNA molecules present a plethora of seconda...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-01-01
|
| Series: | Applied Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-3417/12/3/1543 |
| _version_ | 1827661567863291904 |
|---|---|
| author | Marcin Ryczek Martyna Pluta Leszek Błaszczyk Agnieszka Kiliszek |
| author_facet | Marcin Ryczek Martyna Pluta Leszek Błaszczyk Agnieszka Kiliszek |
| author_sort | Marcin Ryczek |
| collection | DOAJ |
| description | In recent years, it has become clear that RNA molecules are involved in almost all vital cellular processes and pathogenesis of human disorders. The functional diversity of RNA comes from its structural richness. Although composed of only four nucleotides, RNA molecules present a plethora of secondary and tertiary structures critical for intra and intermolecular contacts with other RNAs and ligands (proteins, small metabolites, etc.). In order to fully understand RNA function it is necessary to define its spatial structure. Crystallography, nuclear magnetic resonance and cryogenic electron microscopy have demonstrated considerable success in determining the structures of biologically important RNA molecules. However, these powerful methods require large amounts of sample. Despite their limitations, chemical synthesis and in vitro transcription are usually employed to obtain milligram quantities of RNA for structural studies, delivering simple and effective methods for large-scale production of homogenous samples. The aim of this paper is to provide an overview of methods for large-scale RNA synthesis with emphasis on chemical synthesis and in vitro transcription. We also present our own results of testing the efficiency of these approaches in order to adapt the material acquisition strategy depending on the desired RNA construct. |
| first_indexed | 2024-03-10T00:11:12Z |
| format | Article |
| id | doaj.art-76f70ba36de843f6b0c12cdd7185c77e |
| institution | Directory Open Access Journal |
| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-03-10T00:11:12Z |
| publishDate | 2022-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Applied Sciences |
| spelling | doaj.art-76f70ba36de843f6b0c12cdd7185c77e2023-11-23T15:59:21ZengMDPI AGApplied Sciences2076-34172022-01-01123154310.3390/app12031543Overview of Methods for Large-Scale RNA SynthesisMarcin Ryczek0Martyna Pluta1Leszek Błaszczyk2Agnieszka Kiliszek3Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, PolandInstitute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, PolandInstitute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, PolandInstitute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznań, PolandIn recent years, it has become clear that RNA molecules are involved in almost all vital cellular processes and pathogenesis of human disorders. The functional diversity of RNA comes from its structural richness. Although composed of only four nucleotides, RNA molecules present a plethora of secondary and tertiary structures critical for intra and intermolecular contacts with other RNAs and ligands (proteins, small metabolites, etc.). In order to fully understand RNA function it is necessary to define its spatial structure. Crystallography, nuclear magnetic resonance and cryogenic electron microscopy have demonstrated considerable success in determining the structures of biologically important RNA molecules. However, these powerful methods require large amounts of sample. Despite their limitations, chemical synthesis and in vitro transcription are usually employed to obtain milligram quantities of RNA for structural studies, delivering simple and effective methods for large-scale production of homogenous samples. The aim of this paper is to provide an overview of methods for large-scale RNA synthesis with emphasis on chemical synthesis and in vitro transcription. We also present our own results of testing the efficiency of these approaches in order to adapt the material acquisition strategy depending on the desired RNA construct.https://www.mdpi.com/2076-3417/12/3/1543RNARNA crystallographylarge-scale RNA synthesisribozymes |
| spellingShingle | Marcin Ryczek Martyna Pluta Leszek Błaszczyk Agnieszka Kiliszek Overview of Methods for Large-Scale RNA Synthesis Applied Sciences RNA RNA crystallography large-scale RNA synthesis ribozymes |
| title | Overview of Methods for Large-Scale RNA Synthesis |
| title_full | Overview of Methods for Large-Scale RNA Synthesis |
| title_fullStr | Overview of Methods for Large-Scale RNA Synthesis |
| title_full_unstemmed | Overview of Methods for Large-Scale RNA Synthesis |
| title_short | Overview of Methods for Large-Scale RNA Synthesis |
| title_sort | overview of methods for large scale rna synthesis |
| topic | RNA RNA crystallography large-scale RNA synthesis ribozymes |
| url | https://www.mdpi.com/2076-3417/12/3/1543 |
| work_keys_str_mv | AT marcinryczek overviewofmethodsforlargescalernasynthesis AT martynapluta overviewofmethodsforlargescalernasynthesis AT leszekbłaszczyk overviewofmethodsforlargescalernasynthesis AT agnieszkakiliszek overviewofmethodsforlargescalernasynthesis |