PXD057761-1

PXD057761 is an original dataset announced via ProteomeXchange.

Title	5′ terminal nucleotide determines the immunogenicity of IVT RNAs
Description	In vitro transcription (IVT) is a technology of vital importance that facilitated the production of mRNA therapeutics and drove numerous breakthroughs in RNA biology. T7 polymerase-produced RNAs can begin with either 5′-triphosphate guanosine (5′-pppG) or 5′-triphosphate adenosine (5′-pppA), generating potential agonists for the RIG-I/type I interferon response. While it is established that IVT can yield highly immunogenic double-stranded RNA (dsRNA) via promoterless transcription, the specific contribution of initiating nucleosides to this process has not been previously reported. Our study shows that IVT-derived RNAs containing 5′-pppA are significantly more immunogenic compared with their 5′-pppG counterparts. We observed heightened levels of dsRNAs triggered by IVT with 5′-pppA RNA, activating the RIG-I signaling pathway in cultured cells, as well as in ex vivo and in vivo mouse models where the IFN-β gene was substituted with the mKate2+ reporter. Elevated levels of dsRNA were found in both short 5′-pppA RNAs and full-length mRNAs, including those of COVID-19 vaccines. These findings reveal the unexpected source of IVT RNA immunogenicity, offering valuable insights for both academic research and medical applications of this technology.
HostingRepository	PRIDE
AnnounceDate	2025-02-18
AnnouncementXML	Submission_2025-02-18_08:19:03.845.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jacek Szymanski
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	timsTOF Ultra

Revision	Datetime	Status	ChangeLog Entry
0	2024-11-11 17:39:16	ID requested
⏵ 1	2025-02-18 08:19:04	announced

10.1093/NAR/GKAE1252;

submitter keyword: innate immunity,IVT, dsRNA, RIG-I

Gracjan Michlewski
contact affiliation	International Institute of Molecular and Cell Biology, Laboratory of RNA-Protein Interactions - Dioscuri Centre, Warsaw, Poland
contact email	gmichlewski@iimcb.gov.pl
lab head
Jacek Szymanski
contact affiliation	International Institute of Molecular and Cell Biology in Warsaw, Poland
contact email	jszymanski@iimcb.gov.pl
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/02/PXD057761

PRIDE project URI

• PRIDE
  1. PXD057761
     1. Label: PRIDE project
     2. Name: 5′ terminal nucleotide determines the immunogenicity of IVT RNAs