Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA
Abstract RNA performs a wide range of functions regulated by its structure, dynamics, and often post-transcriptional modifications. While NMR is the leading method for understanding RNA structure and dynamics, it is currently limited by the inability to reduce spectral crowding by efficient segmenta...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2023-12-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-44254-3 |
| _version_ | 1797376888831737856 |
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| author | Raphael Haslecker Vincent V. Pham David Glänzer Christoph Kreutz Theodore Kwaku Dayie Victoria M. D’Souza |
| author_facet | Raphael Haslecker Vincent V. Pham David Glänzer Christoph Kreutz Theodore Kwaku Dayie Victoria M. D’Souza |
| author_sort | Raphael Haslecker |
| collection | DOAJ |
| description | Abstract RNA performs a wide range of functions regulated by its structure, dynamics, and often post-transcriptional modifications. While NMR is the leading method for understanding RNA structure and dynamics, it is currently limited by the inability to reduce spectral crowding by efficient segmental labeling. Furthermore, because of the challenging nature of RNA chemistry, the tools being developed to introduce site-specific modifications are increasingly complex and laborious. Here we use a previously designed Tgo DNA polymerase mutant to present SegModTeX — a versatile, one-pot, copy-and-paste approach to address these challenges. By precise, stepwise construction of a diverse set of RNA molecules, we demonstrate the technique to be superior to RNA polymerase driven and ligation methods owing to its substantially high yield, fidelity, and selectivity. We also show the technique to be useful for incorporating some fluorescent- and a wide range of other probes, which significantly extends the toolbox of RNA biology in general. |
| first_indexed | 2024-03-08T19:45:05Z |
| format | Article |
| id | doaj.art-0eec05fa87c24e6c858477c204ea4afb |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-08T19:45:05Z |
| publishDate | 2023-12-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-0eec05fa87c24e6c858477c204ea4afb2023-12-24T12:23:11ZengNature PortfolioNature Communications2041-17232023-12-0114111210.1038/s41467-023-44254-3Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNARaphael Haslecker0Vincent V. Pham1David Glänzer2Christoph Kreutz3Theodore Kwaku Dayie4Victoria M. D’Souza5Department of Molecular and Cellular Biology, Harvard UniversityDepartment of Molecular and Cellular Biology, Harvard UniversityInstitute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of InnsbruckInstitute of Organic Chemistry and Center for Molecular Biosciences Innsbruck (CMBI), University of InnsbruckDepartment of Chemistry and Biochemistry, University of MarylandDepartment of Molecular and Cellular Biology, Harvard UniversityAbstract RNA performs a wide range of functions regulated by its structure, dynamics, and often post-transcriptional modifications. While NMR is the leading method for understanding RNA structure and dynamics, it is currently limited by the inability to reduce spectral crowding by efficient segmental labeling. Furthermore, because of the challenging nature of RNA chemistry, the tools being developed to introduce site-specific modifications are increasingly complex and laborious. Here we use a previously designed Tgo DNA polymerase mutant to present SegModTeX — a versatile, one-pot, copy-and-paste approach to address these challenges. By precise, stepwise construction of a diverse set of RNA molecules, we demonstrate the technique to be superior to RNA polymerase driven and ligation methods owing to its substantially high yield, fidelity, and selectivity. We also show the technique to be useful for incorporating some fluorescent- and a wide range of other probes, which significantly extends the toolbox of RNA biology in general.https://doi.org/10.1038/s41467-023-44254-3 |
| spellingShingle | Raphael Haslecker Vincent V. Pham David Glänzer Christoph Kreutz Theodore Kwaku Dayie Victoria M. D’Souza Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA Nature Communications |
| title | Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA |
| title_full | Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA |
| title_fullStr | Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA |
| title_full_unstemmed | Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA |
| title_short | Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA |
| title_sort | extending the toolbox for rna biology with segmodtex a polymerase driven method for site specific and segmental labeling of rna |
| url | https://doi.org/10.1038/s41467-023-44254-3 |
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