Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2000 May 15;28(10):E45.
doi: 10.1093/nar/28.10.e45.

Antisense delivery using protamine-oligonucleotide particles

M Junghans  1 , J Kreuter, A Zimmer
Affiliations

Antisense delivery using protamine-oligonucleotide particles

M Junghans et al. Nucleic Acids Res. .

Abstract

Protamine, a polycationic peptide (mol. wt 4000-4500), was evaluated as a potential penetration enhancer for phosphodiester antisense oligonucleotides (ODNs). Unique complexes in the form of nanoparticles were spontaneously formed, which we call 'proticles'. The stability of the particles and the ODNs bound into the proticles was examined in foetal calf serum and cell culture medium. FITC-labelled ODNs bound to protamine showed an increased cellular uptake into human histiocytic lymphoma U 937 cells compared to free ODNs. Proticles significantly decreased cellular growth in a cell proliferation assay using ODNs against the c- myc proto-oncogene.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Hydrodynamic diameters of protamine/ODN particles versus time.
Figure 2
Figure 2
Scanning electron micrograph of 1:1 proticles.
Figure 3
Figure 3
Release of oligonucleotides from proticles in PBS. Free ODN (diamonds), proticles with mass ratios of 5:1 (squares), 2.5:1 (triangles), 1:1 (crosses) and 0.5:1 (double crosses).
Figure 4
Figure 4
Zeta potentials of proticles at different mass ratios protamine/ODNs.
Figure 5
Figure 5
Stability of free and complexed ODNs in cell culture medium versus incubation time. Free ODN (diamonds), proticles with mass ratios of 5:1 (squares), 2.5:1 (trianges), 1:1 (crosses) and 0.5:1 (double crosses).
Figure 6
Figure 6
Laser scanning confocal microscopy of oligonucleotide uptake. U 937 cells were incubated for 4 h at 37°C with (A) free 5′-FITC-labelled phosphodiester oligonucleotides, (B) 1:1 protamine/FODN particles and (C) 2.5:1 protamine/FODN particles. These CLSM pictures are section views showing horizontal and vertical sections of 40 pictures.
Figure 7
Figure 7
Cellular viability after 5 days incubation in a medium control (dot pattern), with unbound and complexed antisense (filled bars), sense (empty bars) and scrambled c-myc ODNs (diagonal lines) or protamine control solution (chessboard pattern). Statistically significant differences (Student’s t-test) between unbound ODNs and proticle preparations (n = 3–6, P < 0.05), as well as between antisense proticles and scrambled proticles (n = 6, P < 0,05) (stars) no significant differences between unbound ODNs and protamine control solution (two stars).
Figure 8
Figure 8
Cellular growth of U 937 cells incubated with medium (diamonds), free antisense c-myc phosphodiester ODNs (squares) and 1:1 (crosses) and 2.5:1 proticles (triangles).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Shi Y., Hutchinson,H.G., Hall,D.J. and Zalewski,A. (1993) Circulation, 88, 1190–1195. - PubMed
    1. Edgington S.M. (1993) Biotechnology, 11, 787–792. - PubMed
    1. Kaddurah-Daouk R., Greene,J.M., Baldwin,A.S.,Jr and Kingston,R.E. (1987) Genes Dev., 1, 347–357. - PubMed
    1. Shi Y., Glynn,J.M., Guilbert,L.J., Cotter,T.G., Bissonnette,R.P. and Green,D.R. (1992) Science, 257, 212–214. - PubMed
    1. Bissonnette R.P., Echeverri,F., Mahboubi,A. and Green,D.R. (1992) Nature, 359, 552–554. - PubMed

Publication types