Abstract
Lipid nanoparticles (LNPs) are established in the biopharmaceutical industry for efficient encapsulation and cytosolic delivery of nucleic acids for potential therapeutics, with several formulations in clinical trials. The advantages of LNPs can also be applied in basic research and discovery with a microfluidic method of preparation now commercially available that allows preparations to be scaled down to quantities appropriate for cell culture. These preparations conserve expensive nucleic acids while maintaining the particle characteristics that have made LNPs successful in later stages of genetic medicine development. Additionally, this method and the resulting LNPs are seamlessly scalable to quantities appropriate for in vivo models and development of nucleic acid therapeutics.
The present work describes the methodology for preparing LNPs loaded with siRNA, mRNA or plasmids using a commercially available microfluidic instrument and an accompanying transfection kit. Guidelines for application to cultured cells in a well-plate format are also provided.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Cullis PR, Hope MJ (2017) Lipid nanoparticle systems for enabling gene therapies. Mol Ther 25:1467–1475. https://doi.org/10.1016/j.ymthe.2017.03.013
Belliveau NM, Huft J, Lin PJ et al (2012) Microfluidic synthesis of highly potent limit-size lipid nanoparticles for in vivo delivery of siRNA. Mol Ther Nucleic Acids 1:e37. https://doi.org/10.1038/mtna.2012.28
Lv H, Zhang S, Wang B et al (2006) Toxicity of cationic lipids and cationic polymers in gene delivery. J Control Release 114:100–109. https://doi.org/10.1016/j.jconrel.2006.04.014
Wu L, Nguyen LH, Zhou K et al (2015) Precise let-7 expression levels balance organ regeneration against tumor suppression. elife 4:e09431. https://doi.org/10.7554/eLife.09431
Yaghi NK, Wei J, Hashimoto Y et al (2017) Immune modulatory nanoparticle therapeutics for intracerebral glioma. Neuro Oncol 19:372–382. https://doi.org/10.1093/neuonc/now198
Ramishetti S, Landesman-Milo D, Peer D (2016) Advances in RNAi therapeutic delivery to leukocytes using lipid nanoparticles. J Drug Target 24:780–786. https://doi.org/10.3109/1061186X.2016.1172587
Weinstein S, Toker IA, Emmanuel R et al (2016) Harnessing RNAi-based nanomedicines for therapeutic gene silencing in B-cell malignancies. Proc Natl Acad Sci U S A 113:E16–E22. https://doi.org/10.1073/pnas.1519273113
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Thomas, A. et al. (2018). Microfluidic Production and Application of Lipid Nanoparticles for Nucleic Acid Transfection. In: Heuck, C., Weinhold, N. (eds) Multiple Myeloma. Methods in Molecular Biology, vol 1792. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-7865-6_14
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7865-6_14
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-7864-9
Online ISBN: 978-1-4939-7865-6
eBook Packages: Springer Protocols