Abstract
The potential of RNAi therapies has been largely impeded by the inherent challenges in the functional delivery of siRNA to cells. Herein, we describe protocols for the synthesis and characterization of novel peptide–siRNA nanoparticles prepared from disulfide-constrained amphipathic peptides complexed with siRNA as promising siRNA delivery vectors. We also describe protocols for the application of these nanoparticles to the in vitro and in vivo delivery of siRNA to lung cells for the functional knockdown of lung proteins.
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
Schwarz DS, Hutvagner G, Du T et al (2003) Asymmetry in the assembly of the RNAi enzyme complex. Cell 115:199–208
Tatiparti K, Sau S, Kashaw SK, Iyer AK (2017) siRNA delivery strategies: a comprehensive review of recent developments. Nanomaterials 7(4):77
Fire A, Xu S, Montgomery MK et al (1998) Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391:806–811
Whitehead KA, Langer R, Anderson DG (2009) Knocking down barriers: advances in siRNA delivery. Nat Rev Drug Discov 8:129
Beierlein JM, Mcnamee LM, Ledley FD (2017) As Technologies for nucleotide therapeutics mature, products emerge. Mol Ther Nucleic Acid 9:379–386
Garber K (2017) Worth the RISC? Nat Biotechnol 35:198
Chakraborty C, Sharma AR, Sharma G et al (2017) Therapeutic miRNA and siRNA: moving from bench to clinic as next generation medicine. Mol Ther Nucleic Acids 8:132–143
Garber K (2018) Alnylam launches era of RNAi drugs. Nat Biotechnol 36:777
Ho W, Zhang X-Q, Xu X (2016) Biomaterials in siRNA delivery: a comprehensive review. Adv Healthc Mater 5:2715–2731
Osborn MF, Khvorova A (2018) Improving siRNA delivery in vivo through lipid conjugation. Nucleic Acid Ther 28:128–136
Juliano RL, Ming X, Nakagawa O (2012) Cellular uptake and intracellular trafficking of antisense and siRNA oligonucleotides. Bioconjug Chem 23:147–157
Wittrup A, Lieberman J (2015) Knocking down disease: a progress report on siRNA therapeutics. Nat Rev Genet 16:543–552
Endoh T, Ohtsuki T (2009) Cellular siRNA delivery using cell-penetrating peptides modified for endosomal escape. Adv Drug Deliv Rev 61:704–709
Tai W, Gao X (2017) Functional peptides for siRNA delivery. Adv Drug Deliv Rev 110–111:157–168
Ahmadzada T, Reid G, McKenzie DR (2018) Fundamentals of siRNA and miRNA therapeutics and a review of targeted nanoparticle delivery systems in breast cancer. Biophys Rev 10:69–86
Welch JJ, Swanekamp RJ, King C et al (2016) Functional delivery of siRNA by disulfide-constrained cyclic amphipathic peptides. ACS Med Chem Lett 7:584–589
Bowerman CJ, Nilsson BL (2010) A reductive trigger for peptide self-assembly and hydrogelation. J Am Chem Soc 132:9526–9527
Mandal D, Nasrolahi Shirazi A, Parang K (2011) Cell-penetrating homochiral cyclic peptides as nuclear-targeting molecular transporters. Angew Chem Int Ed Engl 50:9633–9637
Aitken A, Learmonth M (2009) In: Walker JM (ed) Estimation of disulfide bonds using Ellman’s reagent BT - the protein protocols handbook. Humana Press, Totowa, NJ, pp 1053–1055
O’Nuallain B, Thakur AK, Williams AD et al (2006) Kinetics and thermodynamics of amyloid assembly using a high-performance liquid chromatography-based sedimentation assay. Methods Enzymol 413:34–74
Chan W, White P (1999) Fmoc solid phase peptide synthesis: a practical approach. OUP, Oxford
Han Y, Albericio F, Barany G (1997) Occurrence and minimization of cysteine racemization during stepwise solid-phase peptide Synthesis1,2. J Org Chem 62:4307–4312
Lukszo J, Patterson D, Albericio F, Kates SA (1996) 3-(1-Piperidinyl)alanine formation during the preparation ofC-terminal cysteine peptides with the Fmoc/t-Bu strategy. Lett Pept Sci 3:157–166
Cline DJ, Thorpe C, P Schneider J (2004) General method for facile intramolecular disulfide formation in synthetic peptides. Anal Biochem 335:168–170
Acknowledgments
This work was supported by the National Science Foundation (DMR-1148836) and the National Institutes of Health (R01HL138538, R01HL120521, and EB9903). We thank Karen Bentley and Gayle Schneider of the University of Rochester Medical Center Electron Microscopy Core for assistance with transmission electron microscopy and Jermaine Jenkins of the University of Rochester Medical Center for assistance with dynamic light scattering.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Welch, J.J., Dean, D.A., Nilsson, B.L. (2021). Synthesis and Application of Peptide–siRNA Nanoparticles from Disulfide-Constrained Cyclic Amphipathic Peptides for the Functional Delivery of Therapeutic Oligonucleotides to the Lung. In: Ryadnov, M. (eds) Polypeptide Materials. Methods in Molecular Biology, vol 2208. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0928-6_4
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0928-6_4
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0927-9
Online ISBN: 978-1-0716-0928-6
eBook Packages: Springer Protocols