Weichmann, Franziska and Hett, Robert and Schepers, Aloys and Ito-Kureha, Taku and Flatley, Andrew and Slama, Kaouthar and Hastert, Florian D. and Angstman, Nicholas B. and Cardoso, M. Cristina and Koenig, Julian and Huettelmaier, Stefan and Dieterich, Christoph and Canzar, Stefan and Helm, Mark and Heissmeyer, Vigo and Feederle, Regina and Meister, Gunter (2020) Validation strategies for antibodies targeting modified ribonucleotides. RNA, 26 (10). pp. 1489-1506. ISSN 1355-8382, 1469-9001
Full text not available from this repository. (Request a copy)Abstract
Chemical modifications are found on almost all RNAs and affect their coding and noncoding functions. The identification of m(6)A on mRNA and its important role in gene regulation stimulated the field to investigate whether additional modifications are present on mRNAs. Indeed, modifications including m(1)A, m(5)C, m(7)G, 2'-OMe, and Psi were detected. However, since their abundances are low and tools used for their corroboration are often not well characterized, their physiological relevance remains largely elusive. Antibodies targeting modified nucleotides are often used but have limitations such as low affinity or specificity. Moreover, they are not always well characterized and due to the low abundance of the modification, particularly on mRNAs, generated data sets might resemble noise rather than specific modification patterns. Therefore, it is critical that the affinity and specificity is rigorously tested using complementary approaches. Here, we provide an experimental toolbox that allows for testing antibody performance prior to their use.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | MESSENGER-RNA; RIBOSOMAL-RNA; M(6)A; METHYLATION; PSEUDOURIDYLATION; NUCLEAR; N-6-METHYLADENOSINE; 5-METHYLCYTOSINE; PROTEINS; monoclonal antibodies; affinity; m(6)A; m(5)C; validation; base modifications |
| Subjects: | 500 Science > 570 Life sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I Biology, Preclinical Medicine > Institut für Biochemie, Genetik und Mikrobiologie > Lehrstuhl für Biochemie I > Prof. Dr. Gunter Meister |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 10 Mar 2021 12:47 |
| Last Modified: | 10 Mar 2021 12:47 |
| URI: | https://pred.uni-regensburg.de/id/eprint/43725 |
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