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Rapid and sensitive detection of native glycoRNAs

Helena Hemberger, View ORCID ProfilePeiyuan Chai, Charlotta G. Lebedenko, Reese M. Caldwell, Benson M. George, View ORCID ProfileRyan A. Flynn
doi: https://doi.org/10.1101/2023.02.26.530106
Helena Hemberger
1Stem Cell Program and Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA
2Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA
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Peiyuan Chai
1Stem Cell Program and Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA
2Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA
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  • ORCID record for Peiyuan Chai
Charlotta G. Lebedenko
1Stem Cell Program and Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA
2Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA
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Reese M. Caldwell
1Stem Cell Program and Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA
2Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA
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Benson M. George
1Stem Cell Program and Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA
2Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA
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Ryan A. Flynn
1Stem Cell Program and Division of Hematology/Oncology, Boston Children’s Hospital, Boston, MA, USA
2Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, MA, USA
3Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA
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  • ORCID record for Ryan A. Flynn
  • For correspondence: ryan.flynn@childrens.harvard.edu
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Abstract

Chemical tools enable precise characterization of many biopolymers, including glycoconjugates. Metabolic chemical reporters enabled the discovery of glycoRNAs, however they have certain limitations due the requirement of having living cells to incorporate the modified sugar. Here we develop a periodate oxidation and aldehyde ligation method to detect and characterize native sialoglycoRNAs, termed rPAL. With optimized RNA biochemistry to enhance recovery and analysis of small RNAs, we show rPAL is at least an order of magnitude more sensitive than previous methods for detecting sialoglycoRNAs. These improvements allow rPAL to detect sialoglycoRNA from human clinical samples as demonstrated by defining the abundance and patterns of sialoglycoRNAs from sorted populations of peripheral blood mononuclear cells. The sensitivity, robustness, and flexibility of rPAL will allow greater access towards characterizing glycoRNA biology.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted February 26, 2023.
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Rapid and sensitive detection of native glycoRNAs
Helena Hemberger, Peiyuan Chai, Charlotta G. Lebedenko, Reese M. Caldwell, Benson M. George, Ryan A. Flynn
bioRxiv 2023.02.26.530106; doi: https://doi.org/10.1101/2023.02.26.530106
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Rapid and sensitive detection of native glycoRNAs
Helena Hemberger, Peiyuan Chai, Charlotta G. Lebedenko, Reese M. Caldwell, Benson M. George, Ryan A. Flynn
bioRxiv 2023.02.26.530106; doi: https://doi.org/10.1101/2023.02.26.530106

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