mBACS enhances pseudouridine profiling through efficient and robust chemical conversion
- 1University of Oxford
Abstract
Pseudouridine ({Psi}) is the most abundant RNA modification and regulates RNA stability, splicing, and translation. Utilizing the dual nucleophilic N1 and O2 of {Psi}, we previously developed 2-bromoacrylamide-assisted cyclization sequencing (BACS) for quantitative, single-base {Psi} detection. Here, we screened new dual electrophiles and developed methyl 2-bromoacrylate-assisted cyclization sequencing (mBACS), which achieves higher {Psi} conversion efficiency and lower false-positive rates, enabling more sensitive and robust {Psi} detection. The enhanced sensitivity of mBACS uncovered new {Psi} sites in human tRNAs. mBACS further revealed TRUB1 and PUS10 as the exclusive redundant writers of the conserved tRNA {Psi}55 modification and uncovered crosstalk between {Psi}55 and other tRNA modifications. It also revealed 5-fluorouracil as a pan-pseudouridine synthase inhibitor that induces widespread but site-specific pseudouridylation remodelling. Finally, mBACS supported robust {Psi} profiling from as little as 10 ng of total RNA, establishing it as a sensitive, quantitative, and low-input second-generation platform.
Related
- Cell-resolved pharmacodynamic profiling reveals tissue- and cell-population-specific activity of antisense oligonucleotide chemical conjugates in vivo
- Chemical profiling and multitarget computational screening reveal bioactive phytochemical scaffolds from Echiochilon pauciflorum stocks ex wight
- Integrated non-targeted metabolomics, volatile profiling and targeted quantification characterize chemical changes in Jinhua jasmine tea during the scenting process
- Functional Classification of Zn2+-Dependent Histone Deacetylase Homologs and Variants Using Chemical Profiling
- Insights into the Mechanism of Allele Conversion using a Fluorescent Reporter-Based Compound Heterozygous Allele Conversion Reporter (CHACR) Cell Line Model
BibTeX
@misc{feng2026mbacs,
title = {{mBACS enhances pseudouridine profiling through efficient and robust chemical conversion}},
author = {F. Feng and G. Pisignano and L. Kong and T. Grimes and P. E. Brennan and C. Song},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.05.749584},
url = {https://www.biorxiv.org/content/10.64898/2026.09.05.749584v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - mBACS enhances pseudouridine profiling through efficient and robust chemical conversion
AU - F. Feng
AU - G. Pisignano
AU - L. Kong
AU - T. Grimes
AU - P. E. Brennan
AU - C. Song
PY - 2026
DA - 2026/09/05
PB - bioRxiv
DO - 10.64898/2026.09.05.749584
UR - https://www.biorxiv.org/content/10.64898/2026.09.05.749584v1
AB - Pseudouridine ({Psi}) is the most abundant RNA modification and regulates RNA stability, splicing, and translation. Utilizing the dual nucleophilic N1 and O2 of {Psi}, we previously developed 2-bromoacrylamide-assisted cyclization sequencing (BACS) for quantitative, single-base {Psi} detection. Here, we screened new dual electrophiles and developed methyl 2-bromoacrylate-assisted cyclization sequencing (mBACS), which achieves higher {Psi} conversion efficiency and lower false-positive rates, enabling more sensitive and robust {Psi} detection. The enhanced sensitivity of mBACS uncovered new {Psi} sites in human tRNAs. mBACS further revealed TRUB1 and PUS10 as the exclusive redundant writers of the conserved tRNA {Psi}55 modification and uncovered crosstalk between {Psi}55 and other tRNA modifications. It also revealed 5-fluorouracil as a pan-pseudouridine synthase inhibitor that induces widespread but site-specific pseudouridylation remodelling. Finally, mBACS supported robust {Psi} profiling from as little as 10 ng of total RNA, establishing it as a sensitive, quantitative, and low-input second-generation platform.
ER -