CaltechAUTHORS
  A Caltech Library Service

Physicochemical and Biological Characterization of Targeted, Nucleic Acid-Containing Nanoparticles

Bartlett, Derek W. and Davis, Mark E. (2007) Physicochemical and Biological Characterization of Targeted, Nucleic Acid-Containing Nanoparticles. Bioconjugate Chemistry, 18 (2). pp. 456-468. ISSN 1043-1802. PMCID PMC2517011. doi:10.1021/bc0603539. https://resolver.caltech.edu/CaltechAUTHORS:20170131-080032126

[img] PDF - Accepted Version
See Usage Policy.

4MB

Use this Persistent URL to link to this item: https://resolver.caltech.edu/CaltechAUTHORS:20170131-080032126

Abstract

Nucleic acid-based therapeutics have the potential to provide potent and highly specific treatments for a variety of human ailments. However, systemic delivery continues to be a significant hurdle to success. Multifunctional nanoparticles are being investigated as systemic, nonviral delivery systems, and here, we describe the physicochemical and biological characterization of cyclodextrin-containing polycations (CDP) and their nanoparticles formed with nucleic acids including plasmid DNA (pDNA) and small interfering RNA (siRNA). These polycation/nucleic acid complexes can be tuned by formulation conditions to yield particles with sizes ranging from 60 to 150 nm, ζ potentials from 10 to 30 mV, and molecular weights from ∼7 × 10^7 to 1 × 10^9 g mol^(-1) as determined by light scattering techniques. Inclusion complexes formed between adamantane (AD)-containing molecules and the β-cyclodextrin molecules enable the modular attachment of poly(ethylene glycol) (AD−PEG) conjugates for steric stabilization and targeting ligands (AD−PEG−transferrin) for cell-specific targeting. A 70 nm particle can contain ∼10 000 CDP polymer chains, ∼2000 siRNA molecules, ∼4000 AD−PEG_(5000) molecules, and ∼100 AD−PEG_(5000)−Tf molecules; this represents a significant payload of siRNA and a large ratio of siRNA to targeting ligand (20:1). The particles protect the nucleic acid payload from nuclease degradation, do not aggregate at physiological salt concentrations, and cause minimal erythrocyte aggregation and complement fixation at the concentrations typically used for in vivo application. Uptake of the nucleic acid-containing particles by HeLa cells is measured by flow cytometry and visualized by confocal microscopy. Competitive uptake experiments show that the transferrin-targeted particles display enhanced affinity for the transferrin receptor through avidity effects (multiligand binding). Functional efficacy of the delivered pDNA and siRNA is demonstrated through luciferase reporter protein expression and knockdown, respectively. The analysis of the CDP delivery vehicle provides insights that can be applied to the design of targeted nucleic acid delivery vehicles in general.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/bc0603539DOIArticle
http://pubs.acs.org/doi/abs/10.1021/bc0603539PublisherArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2517011/PubMed CentralArticle
ORCID:
AuthorORCID
Davis, Mark E.0000-0001-8294-1477
Additional Information:© 2007 American Chemical Society. Received 9 November 2006. Published online 28 February 2007. Published in print 1 March 2007. Revised Manuscript Received January 11, 2007. We thank Nicholas Brunelli (California Institute of Technology) for performing the atomic force microscopy imaging and Patrick Koen (California Institute of Technology) for performing the transmission electron microscopy imaging. D.W.B. acknowledges support from a National Science Foundation Graduate Research Fellowship. This publication was made possible by grant number 1 R01 EB004657-01 from the National Institutes of Health (NIH). Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH.
Funders:
Funding AgencyGrant Number
NIH1 R01 EB004657-01
NSF Graduate Research FellowshipUNSPECIFIED
Issue or Number:2
PubMed Central ID:PMC2517011
DOI:10.1021/bc0603539
Record Number:CaltechAUTHORS:20170131-080032126
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20170131-080032126
Official Citation:Physicochemical and Biological Characterization of Targeted, Nucleic Acid-Containing Nanoparticles Derek W. Bartlett and Mark E. Davis Bioconjugate Chemistry 2007 18 (2), 456-468 DOI: 10.1021/bc0603539
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:73844
Collection:CaltechAUTHORS
Deposited By: Ruth Sustaita
Deposited On:31 Jan 2017 17:49
Last Modified:11 Nov 2021 05:22

Repository Staff Only: item control page