Abstract
Nanoparticles consisting of single molecules of DNA condensed with polyethylene glycol-substituted lysine 30-mers efficiently transfect lung epithelium following intrapulmonary administration. Nanoparticles formulated with lysine polymers having different counterions at the time of DNA mixing have distinct geometric shapes: trifluoroacetate or acetate counterions produce ellipsoids or rods, respectively. Based on intracytoplasmic microinjection studies, nanoparticle ellipsoids having a minimum diameter less than the 25 nm nuclear membrane pore efficiently transfect non-dividing cells. This 25 nm size restriction corresponds to a 5.8 kbp plasmid when compacted into spheroids, whereas the 8–11 nm diameter of rod-like particles is smaller than the nuclear pore diameter. In mice, up to 50% of lung cells are transfected after dosing with a rod-like compacted 6.9 kbp lacZ expression plasmid, and correction of the CFTR chloride channel was observed in humans following intranasal administration of a rod-like compacted 8.3 kbp plasmid. To further investigate the potential size and shape limitations of DNA nanoparticles for in vivo lung delivery, reporter gene activity of ellipsoidal and rod-like compacted luciferase plasmids ranging in size between 5.3 and 20.2 kbp was investigated. Equivalent molar reporter gene activities were observed for each formulation, indicating that microinjection size limitations do not apply to the in vivo gene transfer setting.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Liu G, Li D, Pasumarthy MK, Kowalczyk TH, Gedeon C, Hyatt S et al. Nanoparticles of compacted DNA transfect post-mitotic cells. J Biol Chem 2003; 278: 32578–32586.
Kowalczyk TH, Pasumarthy MK, Gedeon C, Hyatt S, Payne JM, Muhammad O et al. Type of polylysine counterion influences morphology and biological function of condensed DNA. Mol Ther 2001; 3: S359 (Abstract 1015).
Ziady AG, Gedeon CR, Miller T, Quan W, Payne JM, Hyatt SL et al. Transfection of airway epithelium by stable PEGylated poly-L-lysine DNA nanoparticles in vivo. Mol Ther 2003; 8: 936–947.
Konstan MW, Davis PB, Wagener JS, Hilliard KA, Stern RC, Milgram LJH et al. Compacted DNA nanoparticles administered to the nasal mucosa of cystic fibrosis subjects are safe and demonstrate partial to complete CFTR reconstitution. Hum Gene Ther 2004; 15: 1255–1269.
Dabauvalle MC, Schulz B, Scheer U, Peters R . Inhibition of nuclear accumulation of karyophilic proteins in living cells by microinjection of the lectin wheat germ agglutinin. Exp Cell Res 1988; 174: 291–296.
Wolff B, Willingham MC, Hanover JA . Nuclear protein import: specificity for transport across the nuclear pore. Exp Cell Res 1988; 178: 318–334.
Lukacs GL, Haggie P, Seksek O, Lechardeur D, Freedman N, Verkman AS . Size-dependent DNA mobility in cytoplasm and nucleus. J Biol Chem 2000; 275: 1625–1629.
Kube D, Davis PB . Intracellular trafficking of non-targeted stabilized molecular conjugates in human airway epithelia cells. Mol Ther 2003; 7: S371 (Abstract 965).
Acknowledgements
We thank Dr Pamela B Davis for review of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Fink, T., Klepcyk, P., Oette, S. et al. Plasmid size up to 20 kbp does not limit effective in vivo lung gene transfer using compacted DNA nanoparticles. Gene Ther 13, 1048–1051 (2006). https://doi.org/10.1038/sj.gt.3302761
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3302761
Keywords
This article is cited by
-
Heat-shrinking DNA nanoparticles for in vivo gene delivery
Gene Therapy (2020)
-
Targeting the photoreceptor cilium for the treatment of retinal diseases
Acta Pharmacologica Sinica (2020)
-
Synergism of wt-p53 and synthetic material in local nano-TAE gene therapy of hepatoma: comparison of four systems and the possible mechanism
BMC Cancer (2019)
-
Intranasal Delivery of pGDNF DNA Nanoparticles Provides Neuroprotection in the Rat 6-Hydroxydopamine Model of Parkinson’s Disease
Molecular Neurobiology (2019)
-
Intranasal administration of plasmid DNA nanoparticles yields successful transfection and expression of a reporter protein in rat brain
Gene Therapy (2014)