Transfection Potency of Lipid Nanoparticles Containing mRNA Depends on Relative Loading Levels
When formulating mRNA into lipid nanoparticles (LNP), various copy numbers of mRNA are encapsulated, leading to a distribution of mRNA loading levels within the LNPs. It is unclear whether the mRNA loading level affects the functional delivery of the message. Here we show that depending on the mRNA loading level, LNPs exhibit distinct mass densities and can be fractionated via ultracentrifugation. Upon fractionation, we investigated if mRNA loading levels influence LNP sizing, lipid composition, and morphology. We further conducted <i>in vitro</i> and <i>in vivo</i> functional delivery of mRNA and found that the LNP fraction with the highest mRNA loading levels was the least transfection competent.
Authors
- J. Cheng (ORCID: https://orcid.org/0000-0002-0496-5542)
- Kenneth W. Harder (ORCID: https://orcid.org/0000-0003-4847-1285)
- Armando Alcázar Magaña (ORCID: https://orcid.org/0000-0002-4731-0247)
- Colin J.D. Ross (ORCID: https://orcid.org/0000-0003-1476-7037)
- Leonard J. Foster (ORCID: https://orcid.org/0000-0001-8551-4817)
- Pieter R. Cullis (ORCID: https://orcid.org/0000-0001-9586-2508)
- Abishek Wadhwa (ORCID: https://orcid.org/0000-0001-6991-1548)
- Kevin Fox (ORCID: https://orcid.org/0009-0008-8316-5674)
- Suiyang Liao (ORCID: https://orcid.org/0000-0003-0705-9112)
- Nuthan Vikas Bathula
- Anna K. Blakney (ORCID: https://orcid.org/0000-0002-5812-9689)
- Alexandra Birkenshaw
- Shuangyu Wang
- Miffy H. Y. Cheng (ORCID: https://orcid.org/0000-0002-0261-4642)
- Chia-Hao Ho (ORCID: https://orcid.org/0000-0003-3670-2680)
- Irafasha C. Casmil (ORCID: https://orcid.org/0000-0003-1890-7930)
Institutions
- University of British Columbia (CA)
- Canada's Michael Smith Genome Sciences Centre (CA)
Publication Details
- Journal
- ACS Applied Materials & Interfaces
- Published
- 2024-12-31
- DOI
- https://doi.org/10.1021/acsami.4c20077
- Citations
- 29
- Primary Topic
- RNA Interference and Gene Delivery
- Type
- article
- Field-Weighted Citation Impact
- 6.78
Funders
- Michael Smith Health Research BC
- Canada Research Chairs
- University of British Columbia
- Canadian Institutes of Health Research
- Natural Sciences and Engineering Research Council of Canada
- Networks of Centres of Excellence of Canada