RNA Modifications & Structure Profiling

Structure profiling and natural modification detection with UltraMarathonRT

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  • Introduction

    RNA Modifications & Structure Profiling with UltraMarathonRT

    UltraMarathonRT (uMRT) is highly sensitive to the modifications deposited to RNA by DMS and SHAPE reagents (such as NAI and 2A3), and thus it has been widely adopted to profile RNA secondary structures using DMS-MaP and SHAPE-MaP. Due to its ultra-high processivity and ability to unwind RNA structures, uMRT can copy any RNA template in a single pass, making it the ideal reverse transcriptase (RT) enzyme for this application. Furthermore, it is the only ultra-processive RT that offers single-molecule resolution. uMRT can enhance the accuracy, coverage, and depth of findings for your RNA structural studies.

    Description

    UltraMarathonRT: Ultimate Tool for High-Resolution RNA Profiling

    1. Ultra-Processivity providing single-molecule resolution
    UltraMarathonRT is the ultimate solution for efficient, full-length cDNA synthesis from long RNA templates. It has been engineered to bind tightly to RNA templates, ensuring minimal dissociation and exceptional end-to-end processivity, making it ideal for RNA structure profiling. Combining the full-length cDNA synthesized by uMRT with nanopore long-read sequencing enables accurate, in-depth structural analysis that provides visibility into distinct structural isoforms which cannot be discerned using traditional, distributive RTs (ref:  Nature Methods (2023) 20, 849–859 and  Nature Protocols (2024) 19, 1835–1865).



    2 Low Error Rates
    UltraMarathonRT offers superior accuracy with lower background error rates compared to traditional reverse transcriptases. By minimizing false-positive mutations, uMRT provides reliable detection of chemical modifications and precise analysis of RNA structural features, ensuring more accurate results in your research.

    3. Decoding Modifications on all four RNA bases in DMS-MaP
    UltraMarathonRT generates unique mutational signatures on DMS modified uracil (U) and guanine (G), making it the only RT enzyme that can accurately detect DMS- induced modifications on all four RNA bases—adenine, cytosine, uracil, and guanine (ref: Nucleic Acids Research (2023) 51, 8744–8757). This expands RNA structural profiling, offering comprehensive nucleotide-level insights, even for previously challenging bases like guanine.

    4. Single-Molecule Resolution
    UltraMarathonRT enables more sensitive and high-resolution single-molecule analysis. This is crucial for detecting correlated modification events, which are used to infer secondary and tertiary structures, as well as dynamic structural states in RNA molecules.

    5. Natural Modification Detection
    UltraMarathonRT is also sensitive to other RNA modifications, making it the first epitranscriptomic tool that detects post-transcriptional modifications (ref:  Journal of Molecular Biology (2023), 435, 168299).



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