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Nucleotide resolution sequencing of N4-acetylcytidine in RNA

  1. Author:
    Thomas,Justin
    Bryson,Keri
    Meier,Jordan

  2. Author Address

    Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, United States., Chemical Biology Laboratory, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Frederick, MD, United States. Electronic address: jordan.meier@nih.gov.,
    1. Year: 2019
    2. Epub Date: 2019 03 12
  2. Journal: Methods in enzymology
    1. 621
    2. Pages: 31-51
  4. Type of Article: Article
  5. ISSN: 0076-6879
  1. Abstract:

    Posttranscriptional modifications of RNA represent an emerging class of regulatory elements in human biology. Improved methods for studying how these elements are controlled and where they occur has the potential to transform our understanding of gene expression in development and disease. Here we describe a chemical method for nucleotide resolution sequencing of N4-acetylcytidine (ac4C), a highly conserved modified nucleobase whose formation is catalyzed by the essential cytidine acetyltransferase enzyme NAT10. This approach enables the sensitive, PCR-amplifiable detection of individual ac4C sites from nanograms of unfractionated cellular RNA. The sensitive and quantitative nature of this assay provides a powerful tool to understand how cytidine acetylation is targeted, profile RNA acetyltransferase dynamics, and validate the sites and stoichiometry of ac4C in novel RNA species. © 2019 Elsevier Inc. All rights reserved.

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External Sources

  1. View Full Text
  2. DOI: 10.1016/bs.mie.2019.02.022
  3. PMID: 31128786
  4. View record in Web of Science®
  5. WOS: 000500713500004
  6. PII : S0076-6879(19)30041-2

Library Notes

  1. Fiscal Year: FY2018-2019
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