ArticleRNA (New York, N.Y.)2022
RNase H-based analysis of synthetic mRNA 5' cap incorporation.
Article in RNA (New York, N.Y.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
17 citing papers in PubMed, 24 citations in OpenAlex.
- A DNA-guided prokaryotic argonaute enables programmable RNA cleavage for sequencing and quality control ofMolecular therapy. Nucleic acids · 2026Article
- Article
- Transcript-independent quantitative analysis of RNA cap structures by LC-MS/MS.Molecular therapy. Advances · 2026Article
- Systematic screening of archaeal MazF homologs reveals Tth-MazF1, a versatile, sequence-specific ribonuclease from Thermococcus thioreducens.Nucleic acids research · 2026Article
- DNA and RNA editing for the therapy of human diseases: current status, challenges, and future prospects.Molecular biomedicine · 2026Review
- Process and analytical strategies for the safe production of mRNA vaccines and therapeutics.Molecular biology reports · 2026Review
- mRNA therapeutics: Transforming medicine through innovation in design, delivery, and disease treatment.Molecular therapy. Nucleic acids · 2025Review
- The strategies and advances of mRNA translation booster.Asian journal of pharmaceutical sciences · 2025Review
- Physicochemical and functional assessment of messenger RNA 5'Cap-end impurities under forced degradation conditions.Molecular therapy. Nucleic acids · 2025Article
- Characterisation and analysis of mRNA critical quality attributes using liquid chromatography based methods.Journal of chromatography. A · 2025Review
- Using nucleolytic toxins as restriction enzymes enables new RNA applications.Nucleic acids research · 2024Article
- Selective Characterization of mRNA 5' End-Capping by DNA Probe-Directed Enrichment with Site-Specific Endoribonucleases.ACS pharmacology & translational science · 2023Article
- Biochemical characterization of mRNA capping enzyme from Faustovirus.RNA (New York, N.Y.) · 2023Article
- Liquid Chromatography Methods for Analysis of mRNA Poly(A) Tail Length and Heterogeneity.Analytical chemistry · 2023Article
- Design and generation of mRNAs encoding conserved regions of SARS-CoV-2 ORF1ab for T cell-mediated immune activation.Future virology · 2023Article
- Application of microfluidic chip electrophoresis for high-throughput nucleic acid fluorescence fragment analysis assays.NAR genomics and bioinformatics · 2023Article
- The Storage and In-Use Stability of mRNA Vaccines and Therapeutics: Not A Cold Case.Journal of pharmaceutical sciences · 2023Review
Corrections and comments
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Authors and funding
8 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Advances in mRNA synthesis and lipid nanoparticles technologies have helped make mRNA therapeutics and vaccines a reality. The 5' cap structure is a crucial modification required to functionalize synthetic mRNA for efficient protein translation in vivo and evasion of cellular innate immune responses. The extent of 5' cap incorporation is one of the critical quality attributes in mRNA manufacturing. RNA cap analysis involves multiple steps: generation of predefined short fragments from the 5' end of the kilobase-long synthetic mRNA molecules using RNase H, a ribozyme or a DNAzyme, enrichment of the 5' cleavage products, and LC-MS intact mass analysis. In this paper, we describe (1) a framework to design site-specific RNA cleavage using RNase H; (2) a method to fluorescently label the RNase H cleavage fragments for more accessible readout methods such as gel electrophoresis or high-throughput capillary electrophoresis; (3) a simplified method for post-RNase H purification using desthiobiotinylated oligonucleotides and streptavidin magnetic beads followed by elution using water. By providing a design framework for RNase H-based RNA 5' cap analysis using less resource-intensive analytical methods, we hope to make RNA cap analysis more accessible to the scientific community.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.