Protocol for in vitro transfection of mRNA-encapsulated lipid nanoparticles
Lipid nanoparticle (LNP) platforms are promising for mRNA therapeutics; however, while highly effective in vivo, commercial mRNA-LNPs show reduced transfection efficiency in vitro under serum-starved conditions, limiting mechanistic studies and cell engineering applications. Here, we present a new standard for in vitro transfection of mRNA-LNPs, including cell culture preparation, mRNA-LNP treatment in complete media, and mRNA expression level quantification. This protocol improves efficiency and reproducibility in in vitro screening and ensures consistency in the assessment of mRNA-LNPs.
Authors
- Eun‐Kyoung Bang (ORCID: https://orcid.org/0000-0002-0339-0740)
- Sangtae Oh (ORCID: https://orcid.org/0009-0006-8368-8868)
- Soyeon Yoo (ORCID: https://orcid.org/0000-0002-5734-2517)
- Haein Lee
- Hyo-wook Gil (ORCID: https://orcid.org/0009-0005-0665-3533)
- Gyochang Keum (ORCID: https://orcid.org/0000-0001-8767-8022)
- Kounghwa Youn (ORCID: https://orcid.org/0009-0008-9022-2008)
- Ha Young Park (ORCID: https://orcid.org/0000-0002-7192-2374)
- In Sung Hwang (ORCID: https://orcid.org/0000-0001-5572-8670)
Institutions
- Sogang University (KR)
- Korea University (KR)
- Kyung Hee University (KR)
- Korea University (JP)
- Korea Institute of Science and Technology (KR)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-10-28
- DOI
- https://doi.org/10.1016/j.xpro.2025.104159
- Primary Topic
- RNA Interference and Gene Delivery
- Type
- article
- Field-Weighted Citation Impact
- 0.00