Rapid and sensitive detection of native glycoRNAs

• DOI: 10.1101/2023.02.26.530106
• Publication Date: 2023-02-27T04:10:12Z
• Authors: Helena Hemberger; Peiyuan Chai; Charlotta G. Lebe...; Reese M. Caldwell; Benson M. George; Ryan A. Flynn
• Keywords: 0303 health sciences; 03 medical and health sciences

AbstractChemical 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.