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Ultra-long Read Sequencing for Whole Genomic DNA Analysis
Published on: March 15, 2019
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Mapping human pre-rRNA processing and modification at single nucleotide resolution using long read nanopore
Stefan Pastore1, Ludivine Wacheul2, Lioba Lehmann3
1Institute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg-University Mainz, Mainz, Germany.
Nature Communications
|March 31, 2026
Summary
NanoRibolyzer, a nanopore sequencing method, precisely identifies and quantifies precursor rRNA (pre-rRNA) molecules and their modifications. This advance offers new insights into ribosome biogenesis and RNA processing pathways.
Area of Science:
- Molecular Biology
- Genomics
- Biochemistry
Background:
- Ribosome biogenesis is crucial for cell function, involving complex precursor rRNA (pre-rRNA) processing.
- Traditional methods for studying pre-rRNA processing lack the resolution needed for detailed analysis.
Purpose of the Study:
- To introduce NanoRibolyzer, a novel nanopore-based long-read sequencing approach.
- To enable ab initio identification, quantification, and modification mapping of pre-rRNAs.
- To provide high-resolution insights into rRNA processing and modification dynamics.
Main Methods:
- Nanopore-based long-read sequencing (NanoRibolyzer).
- Supervised and unsupervised mapping for pre-rRNA identification.
- Cell fractionation for spatial deconvolution of processing.
- Analysis of exoRNase activity and RNA modification mapping (pseudouridine).
Main Results:
- Detection of known and novel pre-rRNAs with single-nucleotide resolution.
- Delineation of rRNA cleavage events and spatial separation of nuclear/cytoplasmic processing.
- Characterization of exoRNase activity and identification of condition-specific processing fingerprints.
- Pseudouridine mapping revealed extensive modification of primary transcripts and hypomodification of aberrant intermediates.
Conclusions:
- NanoRibolyzer significantly advances the study of ribosome biogenesis by providing unprecedented resolution.
- The method reveals novel insights into rRNA processing pathways, RNA modifications, and the functions of ribosome assembly factors.
- NanoRibolyzer has potential for biomarker discovery in RNA processing defects.
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