Insight into mechanisms of cellular uptake of lipid nanoparticles and intracellular release of small RNAs

  • Pharm Res. 2014 Oct;31(10):2685-95. doi: 10.1007/s11095-014-1366-7.
Bo Yu  1 ,  Xinmei Wang ,  Chenguang Zhou ,  Lesheng Teng ,  Wei Ren ,  Zhaogang Yang ,  Chih-Hsin Shih ,  Tianyou Wang ,  Robert J Lee ,  Suoqin Tang ,  L James Lee
Affiliations
  • 1. Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, USA.
PMID: 24740244 DOI: 10.1007/s11095-014-1366-7
Abstract

Purpose: Understanding mechanisms of cellular uptake and intracellular release would enable better design of nanocarriers for delivery of nucleic acids such as siRNA and MicroRNA (miRNA).

Method: In this study, we investigated cellular pharmacokinetics of siRNA by co-encapsulating fluorescently labeled siRNA and molecular beacon (MB) in four different formulations of cationic lipid nanoparticles (LNPs). A miRNA mimic was also used as a probe for investigating cellular pharmacokinetics, which correlated well with RNAi activities.

Results: We tried to find the best LNP formulation based on the combination of DOTMA and DODMA. When the DOTMA/DODMA ratio was at 5/40, the LNP containing a luciferase siRNA produced the highest gene silencing activity. The superior potency of DOTMA/DODMA could be attributed to higher uptake and improved ability to facilitate siRNA release from endosomes subsequent to uptake.

Conclusions: Our findings may provide new insights into RNAi transfection pathways and have implications on cationic LNP design.

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