Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA
Open accessAbstract
Lipid nanoparticles (LNP) are the leading systems for in vivo delivery of small interfering RNA (siRNA) for therapeutic applications. Formulation of LNP siRNA systems requires rapid mixing of solutions containing cationic lipid with solutions containing siRNA. Current formulation procedures employ macroscopic mixing processes to produce systems 70-nm diameter or larger that have variable siRNA encapsulation efficiency, homogeneity, and reproducibility. Here, we show that microfluidic mixing techniques, which permit millisecond mixing at the nanoliter scale, can reproducibly generate limit size LNP siRNA systems 20 nm and larger with essentially complete encapsulation of siRNA over a wide range of conditions with polydispersity indexes as low as 0.02. Optimized LNP siRNA systems produced by microfluidic mixing achieved 50% target gene silencing in hepatocytes at a dose level of 10 µg/kg siRNA in mice. We anticipate that microfluidic mixing, a precisely controlled and readily scalable technique, will become the preferred method for formulation of LNP siRNA delivery systems.
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The LNP delivery systems used here contain the ionizable cationic lipid DMAP-BLP, a lipid that exhibits optimized bilayer destabilizing and pKa properties leading to highly potent gene silencing in hepatocytes following IV administration similar to "gold standard" lipids such as DLinMC3-DMA.
“With respect to the mechanism of action, the LNP delivery systems used here contain the ionizable cationic lipid DMAP-BLP, a lipid that exhibits optimized bilayer destabilizing and pKa properties leading to highly potent gene silencing in hepatocytes following IV administration (Supplementary Data) that is similar to “gold standard” lipids such as DLinMC3-DMA.8”
From Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain (2013) · cites this paper as [8]
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Experimental — accuracy not yet measuredLNP systems with this lipid composition can silence target genes in hepatocytes following IV injection at dose levels as low as 0.01 mg siRNA/kg body weight in an apolipoprotein E (ApoE)-dependent fashion.
“Previous work has shown that LNP systems with this lipid composition can silence target genes in hepatocytes following IV injection at dose levels as low as 0.01 mg siRNA/kg body weight8,9 in an apolipoprotein E (ApoE)–dependent fashion.10”
From Lipid Nanoparticle Delivery of siRNA to Silence Neuronal Gene Expression in the Brain (2013) · cites this paper as [8]
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- Authors
- Belliveau, N. M., Huft, J., Lin, P. J., Chen, S., Leung, A. K., Leaver, T. J., Wild, A. W., Lee, J. B., Taylor, R. J., Tam, Y. K., Hansen, C. L., & Cullis, P. R.
- Affiliations
- University of British Columbia (CA)
- Precision Nanosystems (Canada) (CA)
- Corresponding author
- Cullis, Pieter R
- Venue
- Molecular Therapy - Nucleic Acids
- Publisher
- Elsevier BV
- Published
- 2012
- Volume
- 1
- Pages
- e37
- ISSN
- 2162-2531 (ISSN-L)
- PMC
- PMC3442367
- OpenAlex
- W2125470506
- Cited by
- 685
- References
- 4
- Citations by year
- 2026: 49 · 2025: 85 · 2024: 91 · 2023: 84 · 2022: 67 · 2021: 60 · 2020: 51 · 2019: 46 · 2018: 29 · 2017: 29 · 2016: 37 · 2015: 21 · 2014: 20 · 2013: 12 · 2012: 2
- FWCI
- 7.08
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Molecular Therapy — Nucleic Acids (publishedVersion, cc-by-nc-nd)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by-nc-nd)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Health Sciences › Medicine › Pulmonary and Respiratory Medicine › Inhalation and Respiratory Drug Delivery
- Keywords
- lipid nanoparticles, siRNA delivery, microfluidic mixing, cationic lipids, nanoparticle size control, polydispersity, gene silencing, hepatocytes
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