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Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA
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 m...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2012
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3442367/ https://www.ncbi.nlm.nih.gov/pubmed/23344179 http://dx.doi.org/10.1038/mtna.2012.28 |
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author | Belliveau, Nathan M Huft, Jens Lin, Paulo JC Chen, Sam Leung, Alex KK Leaver, Timothy J Wild, Andre W Lee, Justin B Taylor, Robert J Tam, Ying K Hansen, Carl L Cullis, Pieter R |
author_facet | Belliveau, Nathan M Huft, Jens Lin, Paulo JC Chen, Sam Leung, Alex KK Leaver, Timothy J Wild, Andre W Lee, Justin B Taylor, Robert J Tam, Ying K Hansen, Carl L Cullis, Pieter R |
author_sort | Belliveau, Nathan M |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-3442367 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2012 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-34423672012-09-14 Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA Belliveau, Nathan M Huft, Jens Lin, Paulo JC Chen, Sam Leung, Alex KK Leaver, Timothy J Wild, Andre W Lee, Justin B Taylor, Robert J Tam, Ying K Hansen, Carl L Cullis, Pieter R Mol Ther Nucleic Acids Original Article 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. Nature Publishing Group 2012-08 2012-08-14 /pmc/articles/PMC3442367/ /pubmed/23344179 http://dx.doi.org/10.1038/mtna.2012.28 Text en Copyright © 2012 American Society of Gene & Cell Therapy http://creativecommons.org/licenses/by-nc-nd/3.0/ Molecular Therapy-Nucleic Acids is an open-access journal published by Nature Publishing Group. This work is licensed under the Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/ |
spellingShingle | Original Article Belliveau, Nathan M Huft, Jens Lin, Paulo JC Chen, Sam Leung, Alex KK Leaver, Timothy J Wild, Andre W Lee, Justin B Taylor, Robert J Tam, Ying K Hansen, Carl L Cullis, Pieter R Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA |
title | Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA |
title_full | Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA |
title_fullStr | Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA |
title_full_unstemmed | Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA |
title_short | Microfluidic Synthesis of Highly Potent Limit-size Lipid Nanoparticles for In Vivo Delivery of siRNA |
title_sort | microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of sirna |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3442367/ https://www.ncbi.nlm.nih.gov/pubmed/23344179 http://dx.doi.org/10.1038/mtna.2012.28 |
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