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PXD033334-1

PXD033334 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleHuman RNase 4 Improves mRNA Sequence Characterization by LC-MS/MS.
DescriptionWith the rapid growth of synthetic messenger RNA (mRNA)-based therapeutics and vaccines, the development of analytical tools for characterization of long, complex RNAs has become essential. Tandem liquid chromatography-mass spectrometry (LC-MS/MS) permits direct assessment of the mRNA primary sequence and modifications thereof without conversion to cDNA or amplification. It relies upon digestion of mRNA with site-specific endoribonucleases to generate pools of short oligonucleotides that are then amenable to MS-based sequence analysis. Here, we showed that the uridine-specific human endoribonuclease hRNase 4 improves mRNA sequence coverage, in comparison with the benchmark enzyme RNase T1, by producing a large population of uniquely mappable cleavage products. Furthermore, we deployed hRNase 4 to characterize mRNAs fully substituted with 1-methylpseudouridine (m1Ψ) or 5-methoxyuridine (mo5U), as well as mRNAs selectively depleted of uridine–two key strategies to reduce synthetic mRNA immunogenicity. Lastly, we demonstrated that hRNase 4 enables direct assessment of the 5′ cap incorporation in in vitro synthesized mRNA. Collectively, this study highlights the power of hRNase 4 to interrogate mRNA sequence, identity, and modifications by LC-MS/MS.
HostingRepositoryPRIDE
AnnounceDate2022-07-26
AnnouncementXMLSubmission_2022-07-26_11:08:26.935.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterEric Wolf
SpeciesList scientific name: synthetic RNA; NCBI TaxID: 2086595;
ModificationListmethylated residue
InstrumentQ Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-04-21 05:55:42ID requested
12022-07-26 11:08:27announced
22023-11-14 08:55:39announced2023-11-14: Updated project metadata.
Publication List
10.1093/NAR/GKAC632;
Keyword List
submitter keyword: mRNA
Human RNase4;
Contact List
Ivan R. Correa Jr.
contact affiliationNew England Biolabs, Inc, 240 County Rd, Ipswich, MA, 01938, USA.
contact emailcorrea@neb.com
lab head
Eric Wolf
contact affiliationNew England Biolabs
contact emailewolf@neb.com
dataset submitter
Full Dataset Link List
Dataset FTP location
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PRIDE project URI
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