ISIS Neutron and Muon Source Data Journal

RNA modification via novel fluorescent base analogues: Effect on the structure of RNA-containing lipid nanoparticles and their sub-populations

Abstract: Nucleic acid-based therapies are a rapidly expanding field. To study delivery and performance of nucleic acids in cells, they must often be fluorescently labelled, usually by attaching large fluorescent dyes. However, this can significantly perturb the nucleic acid?s biological functions and structural integrity. To address this, the Wilhelmsson group has developed chemically modified nucleic acid building blocks which are inherently fluorescent, so-called ?fluorescent base analogues?. These can be efficiently incorporated into RNA which closely mimics the behaviour of the native RNA, and can be translated to proteins within cells. LNPs are enormously successful mRNA delivery vectors, but relationships between their structure and function are not yet well understood. To leverage the benefits of fluorescent base analogue labeled RNA to explore critical questions in the field (e.g. how cargo is released from LNPs), the effect of incorporating these novel labelled RNAs on LNP structure needs to be validated. In this experiment, we characterise the structures of LNPs loaded with fluorescent base analogue labeled RNA, and compare them to LNPs loaded with unlabeled and conventionally labeled RNA. We will also characterise sub-populations within these LNP formulations, separated using AF4. The results will deepen our understanding of the impact of fluorescent labelling strategies on LNP architecture, and mark a pivotal step forward in correlating LNP structure to performance.

Principal Investigator: Dr Margaret Holme
Experimenter: Professor Marcus Wilhelmsson
Experimenter: Ms Alma Karlsson
Local Contact: Dr James Doutch
Experimenter: Dr Hanna Barriga
Experimenter: Dr Jennifer Gilbert
Experimenter: Ms Anna-Maria Börjesdotter
Experimenter: Miss Anna Johanna Josefine Järvdalen

DOI: 10.5286/ISIS.E.RB2410726

ISIS Experiment Number: RB2410726

Publisher: STFC ISIS Neutron and Muon Source

Data format: RAW/Nexus
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Data Citation

The recommended format for citing this dataset in a research publication is as:
[author], [date], [title], [publisher], [doi]

For Example:
Dr Margaret Holme et al; (2025): RNA modification via novel fluorescent base analogues: Effect on the structure of RNA-containing lipid nanoparticles and their sub-populations, STFC ISIS Neutron and Muon Source, https://doi.org/10.5286/ISIS.E.RB2410726

Data is released under the CC-BY-4.0 license.