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Weichmann, F.* ; Hett, R.* ; Schepers, A. ; Ito-Kureha, T.* ; Flatley, A. ; Slama, K.* ; Hastert, F.D.* ; Angstman, N.B.* ; Cardoso, M.C.* ; König, J.* ; Hüttelmaier, S.* ; Dieterich, C.* ; Canzar, S.* ; Helm, M.* ; Heissmeyer, V. ; Feederle, R. ; Meister, G.*

Validation strategies for antibodies targeting modified ribonucleotides.

RNA 26, 1489-1506 (2020)
Postprint DOI
Open Access Green
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.
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Publication type Article: Journal article
Document type Scientific Article
Corresponding Author
Keywords Monoclonal Antibodies ; Affinity ; M(6)a ; M(5)c ; Validation ; Base Modifications; Messenger-rna; Ribosomal-rna; M(6)a; Methylation; Pseudouridylation; Nuclear; N-6-methyladenosine; 5-methylcytosine; Proteins
ISSN (print) / ISBN 1355-8382
e-ISSN 1469-9001
Journal RNA
Quellenangaben Volume: 26, Issue: 10, Pages: 1489-1506 Article Number: , Supplement: ,
Publisher Cold Spring Harbor Laboratory Press
Publishing Place 1 Bungtown Rd, Cold Spring Harbor, Ny 11724 Usa
Non-patent literature Publications
Reviewing status Peer reviewed
Institute(s) CF Monoclonal Antibodies (CF-MAB)
Abteilung für Molekulare Immunregulation (AMIR)