Small-molecule affinity capture of DNA/RNA quadruplexes and their identification in vitro and in vivo through the G4RP protocol
| Autor: | Sunny Y. Yang, David Monchaud, Isaline Renard, Apolline Roux, Pauline Lejault, Marc Pirrotta, Judy M. Y. Wong, Michael Grandmougin |
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| Přispěvatelé: | Institut de Chimie Moléculaire de l'Université de Bourgogne [Dijon] (ICMUB), Centre National de la Recherche Scientifique (CNRS)-Université de Bourgogne (UB)-Institut de Chimie du CNRS (INC) |
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Computational biology
Biology G-quadruplex Ligands Transcriptome 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Chemical Biology and Nucleic Acid Chemistry Gene expression Genetics Humans Biotinylation [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology 030304 developmental biology 0303 health sciences Genome Human Reverse Transcriptase Polymerase Chain Reaction RNA [SDV.BBM.BM]Life Sciences [q-bio]/Biochemistry Molecular Biology/Molecular biology DNA G-Quadruplexes chemistry Nucleic acid MCF-7 Cells Human genome 030217 neurology & neurosurgery |
| Zdroj: | Nucleic Acids Research Nucleic Acids Research, Oxford University Press, 2019, 47 (11), pp.5502-5510. ⟨10.1093/nar/gkz215⟩ |
| ISSN: | 0305-1048 1362-4962 |
| DOI: | 10.1093/nar/gkz215⟩ |
| Popis: | International audience; Guanine-rich DNA and RNA sequences can fold into higher-order structures known as G-quadruplexes (or G4-DNA and G4-RNA, respectively). The prevalence of the G4 landscapes in the human genome, transcriptome and ncRNAome (non-coding RNA), collectively known as G4ome, is strongly suggestive of biological relevance at multiple levels (gene expression , replication). Small-molecules can be used to track G4s in living cells for the functional characterization of G4s in both normal and disease-associated changes in cell biology. Here, we describe biotinylated biomimetic ligands referred to as Bio-TASQ and their use as molecular tools that allow for isolating G4s through affinity pull-down protocols. We demonstrate the general applicability of the method by purifying biologically relevant G4s from nucleic acid mixtures in vitro and from human cells through the G4RP-RT-qPCR protocol. Overall, the results presented here represent a step towards the optimization of G4-RNAs identification, a key step in studying G4s in cell biology and human diseases. |
| Databáze: | OpenAIRE |
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