Abstract
Aptamers selected against cell surface receptors represent a unique set of ligands that can be used to target nanoparticles and other therapeutics to specific cell types. Here, we describe a method for using aptamers to deliver stable nucleic acid lipid particles (SNALPs) encapsulating small interfering RNA (siRNA) to cells in vitro. Using this method, we have demonstrated the ability of aptamer-conjugated SNALPs to achieve target-specific delivery and siRNA-mediated knockdown of a gene of interest. We also describe methods to characterize SNALP size, siRNA encapsulation efficiency, and aptamer conjugation efficiency.
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
Semple SC, Akinc A, Chen J et al (2010) Rational design of cationic lipids for siRNA delivery. Nat Biotechnol 28:172–176
Heyes J, Palmer L, Bremner K et al (2005) Cationic lipid saturation influences intracellular delivery of encapsulated nucleic acids. J Control Release 107:276–287
Heyes J, Hall K, Tailor V et al (2006) Synthesis and characterization of novel poly(ethylene glycol)-lipid conjugates suitable for use in drug delivery. J Control Release 112:280–290
Lammers T, Kiessling F, Hennink WE et al (2011) Drug targeting to tumors: principles, pitfalls and (pre-) clinical progress. J Control Release
Davis ME, Zuckerman JE, Choi CHJ et al (2010) Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles. Nature 464:1067–1070
Davis ME (2009) The first targeted delivery of siRNA in humans via a self-assembling, cyclodextrin polymer-based nanoparticle: from concept to clinic. Mol Pharm 6:659–668
Akinc A, Querbes W, De S et al (2010) Targeted delivery of RNAi therapeutics with endogenous and exogenous ligand-based mechanisms. Mol Ther 18:1357–1364
Sato Y, Murase K, Kato J et al (2008) Resolution of liver cirrhosis using vitamin A-coupled liposomes to deliver siRNA against a collagen-specific chaperone. Nat Biotechnol 26:431–442
Tam Y, Chen S, Cullis P (2013) Advances in lipid nanoparticles for siRNA delivery. Pharmaceutics 5:498–507
Tam YYC, Chen S, Zaifman J et al (2013) Small molecule ligands for enhanced intracellular delivery of lipid nanoparticle formulations of siRNA. Nanomedicine 9:665–674
Wilner SE, Wengerter B, Maier K et al (2012) An RNA alternative to human transferrin: a new tool for targeting human cells. Mol Ther Nucleic Acids 1, e21
Ni X, Zhang Y, Ribas J et al (2011) Prostate-targeted radiosensitization via aptamer-shRNA chimeras in human tumor xenografts. J Clin Invest 121:2383–2390
Zhou J, Swiderski P, Li H et al (2009) Selection, characterization and application of new RNA HIV gp 120 aptamers for facile delivery of Dicer substrate siRNAs into HIV infected cells. Nucleic Acids Res 37:3094–3109
Dhar S, Gu FX, Langer R et al (2008) Targeted delivery of cisplatin to prostate cancer cells by aptamer functionalized Pt(IV) prodrug-PLGA-PEG nanoparticles. Proc Natl Acad Sci 105:17356–17361
Zhou J, Rossi JJ (2014) Cell-type-specific, aptamer-functionalized agents for targeted disease therapy. Mol Ther Nucleic Acids 3, e169
Reulen SWA, Brusselaars WWT, Langereis S et al (2007) Protein-liposome conjugates using cysteine-lipids and native chemical ligation. Bioconjug Chem 18:590–596
Leung AKK, Hafez IM, Baoukina S et al (2012) Lipid nanoparticles containing siRNA synthesized by microfluidic mixing exhibit an electron-dense nanostructured core. J Phys Chem C 116:18440–18450
Jayaraman M, Ansell SM, Mui BL (2012) Maximizing the potency of siRNA lipid nanoparticles for hepatic gene silencing in vivo. Angew Chem Int Ed 51(34):8529–8533
Jeffs LB, Palmer LR, Ambegia EG et al (2005) A scalable, extrusion-free method for efficient liposomal encapsulation of plasmid DNA. Pharm Res 22:362–372
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Wilner, S.E., Levy, M. (2016). Synthesis and Characterization of Aptamer-Targeted SNALPs for the Delivery of siRNA. In: Mayer, G. (eds) Nucleic Acid Aptamers. Methods in Molecular Biology, vol 1380. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3197-2_18
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3197-2_18
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3196-5
Online ISBN: 978-1-4939-3197-2
eBook Packages: Springer Protocols