Skip to main content
SpringerLink
Log in

Cellular Internalization of Oligodeoxynucleotides

  • Chapter
Gene Therapeutics

Abstract

Although the addition of oligodeoxynucleotides to cell cultures has been demonstrated to effectively inhibit intracellular gene expression, such findings have been in contrast to the prevailing view that cells are impermeable to negatively charged large molecules. With this in mind, several groups have attempted to modify oligos in order to circumvent the problem of negative charge. Thus, methylphosphonates are ionically neutral, and poly-L-lysine has been linked to oligos to supply a strong positive charge. Others have made various oligonucleotide-protein conjugates to aid internalization, while the benefits of oligo encapsulation into liposomes have also been explored. During the last several years, it has become clear that even highly negatively charged oligonucleotides (i.e., phosphodiesters and phosphorothioates) are actively internalized by cells in culture. Furthermore, it has become equally apparent that backbone or pendant oligonucleotide modifications can alter olionucleotide uptake characteristics, although not always in the way intended. In this chapter, we will review the current state of knowledge concerning internalization pathways for modified and unmodified oligonucleotides and how these pathways may affect the intracellular trafficking of oligonucleotides.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 39.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 54.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • Akhtar S, Basu S, Wickstrom E, Juliano RL (1991): Interactions of antisense DNA oligonucleotide analogs with phospholipid membranes (liposomes). Nucleic Acids Research 19: 5551–5559

    Article  PubMed  CAS  Google Scholar 

  • Bennett RM, Davis J, Campbell S, Portnoff S (1983): Lactoferrin binds to cell membrane DNA. J Clin Invest 71: 611–618

    Article  PubMed  CAS  Google Scholar 

  • Bennett RM, Gabor GT, Merritt MM (1985): DNA binding to human leukocytes. J Clin Invest 76: 2182–2190

    Article  PubMed  CAS  Google Scholar 

  • Bennett RM, Kotzin BL, Merritt MJ (1987): DNA receptor dysfunction in systemic lupus erythematosus and kindred disorders. J Exp Med 166: 850–863

    Article  PubMed  CAS  Google Scholar 

  • Bennett RM, Peller JS, Merritt MM (1986): Defective DNA-receptor function in systemic lupus erythematosus and related diseases: evidence for an autoantibody influencing cell physiology. The Lancet Jan 25: 186–188

    Article  Google Scholar 

  • Boutorin AS, Gus’kova, LV, Ivanova EM, Kobetz ND, Zarytova VF, Ryte AS, Yurchenko LV, Vlassov VV (1989): Synthesis of alkylating oligonucleotide derivatives containing cholesterol of phenazinium residues at their 3-terminus and their interaction with DNA within mammalian cells. FEBS Lett 254: 129–132

    Article  PubMed  CAS  Google Scholar 

  • Chin DJ, Green GA, Zon G, Szoka FCJ, Straubinger RM (1990): Rapid nuclear accumulation of injected oligodeoxyribonucleotides. New Biologist 2: 1091–1100

    PubMed  CAS  Google Scholar 

  • Chu C-J, Dijkstra J, Lai M-Z, Hong K, Szoka FC (1990): Efficiency of cytoplasmic delivery by pH-sensitive liposomes to cells in culture. Pharmaceutical Research 7: 824–834

    Article  PubMed  CAS  Google Scholar 

  • Dautry-Varsat A, Ciechanover A, Lodish HF (1983): pH and the recycling of transferrin during receptor-mediated endocytosis. Proc Natl Acad Sci USA 80: 2258–2262

    Google Scholar 

  • Gabor G, Bennett RM (1984): Biotin-labelled DNA: A novel approach for the recognition of a DNA binding site on cell membrane. Biochem Biophys Res Commun 122: 1034–1039

    Article  PubMed  CAS  Google Scholar 

  • Geselowitz DA, Neckers L (1992): Analysis of oligonucleotide binding, internalization and intracellular trafficking utilizing a novel radiolabeled crosslinker. Antisense Res Dev (In Press)

    Google Scholar 

  • Harel-Bellan A, Ferris DK, Vinocour M, Holt JT, FarrarWL (1988): Specific inhibition of c-myc protein biosynthesis using an antisense synthetic deoxy-oligonucleotide in human lymphocytes. J Immunology 140: 2431–2435

    CAS  Google Scholar 

  • Harewood K, Pape K, Gabel C, Suleske R, Cunningham A (1991): Cellular uptake and localization of fluorescein-labelled, 15 mer phosphorothioate and phosphodiester oligonucleotides. J Cell Biochem 15D: 35

    Google Scholar 

  • Hefeneider SH, Bennett RM, Pham TQ, Cornell K, McCoy SL, Heinrich MC (1990): Identification of a cell-surface DNA receptor and its association with systemic lupus erythematosus. J Invest Dermatol 94: 79S–84S

    Article  PubMed  CAS  Google Scholar 

  • Krangel MS (1987): Endocytosis and recycling of the T3-T cell receptor complex. The role of T3 phosphorylation. J Exp Med 165: 1141–1159

    Article  PubMed  CAS  Google Scholar 

  • Kulka M, Smith CC, Aurelian L, Fishelevich R, Meade K, Miller P, Tso POP (1989): Site specificity of the inhibitory effects of oligo(nucleoside methylphosphonate)s complementary to the acceptor splice junction of herpes simplex virus type 1 immediate early mRNA 4. Proc Natl Acad Sci USA 86: 6868–6872

    Article  PubMed  CAS  Google Scholar 

  • Lemaitre M, Bayard B, Lebleu B (1987): Specific antiviral activity of a poly(L-lysine)-conjugated oligodeoxyribonucleotide sequence complementary to vesicular stomatitis virus N protein mRNA initiation site. Proc Natl Acad Sci USA 84: 648–652

    CAS  Google Scholar 

  • Leonetti J-P, Degols G, Lebleu B (1990): Biological Activity of Oligonucle- otide-Poly (L-lysine) Conjugates: Mechanism of Cell Uptake. Bioconj Chem 1: 149–153

    Article  CAS  Google Scholar 

  • Leonetti JP, Machy P, Degols G, Lebleu B, Leserman L (1990): Antibodytargeted liposomes containing oligodeoxynucleotides complementary to viral RNA selectively inhibit viral replication. Proc Natl Acad Sci USA 87: 2448–2451

    Article  PubMed  CAS  Google Scholar 

  • Leonetti JP, Mechti N, Degols G, Gagnor C, Lebleu B (1991): Intracellular distribution of microinjected antisense oligonucleotides. Proc Natl Acad Sci USA 88: 2702–2706

    Article  PubMed  CAS  Google Scholar 

  • Letsinger RL, Zhang GR, Sun DK, Ikeuchi T, Sarin PS (1989): Cholesteryl-conjugated oligonucleotides: synthesis, properties and activity as inhibitors of replication of human immunodeficiency virus in cell culture. Proc Natl Acad Sci USA 86: 6553–6556

    Article  PubMed  CAS  Google Scholar 

  • Loke SL, Stein CA, Zhang XH, Mori K, Nakanishi M, Subasinghe C, Cohen J S, Neckers LM (1989): Chacterization of oligonucleotide transport into living cells. Proc Natl Acad Sci USA 86: 3474–3478

    Article  PubMed  CAS  Google Scholar 

  • Machy P, Truneh A, Gennaro D, Hoffstein S (1987): Major histocompatibility complex class I molecules internalized via coated pits in T lymphocytes. Nature (London) 328: 724–726

    Article  CAS  Google Scholar 

  • Milhaud PG, Maehy P, Lebleu B, Leserman L (1989): Antibody targeted liposomes containing poly (rl). poly(rC) exert a specific antiviral and toxic effect on cells primed with interferons alpha, beta or gamma. Biochim Biophys Acta 987: 15–20

    Article  PubMed  CAS  Google Scholar 

  • Miller PS, McParland KB, Jayaraman K, Ts’o POP (1981): Biochemical and biological effects of nonionic nucleic acid methylphosphonates. Biochemistry 20: 1874–1880

    Article  PubMed  CAS  Google Scholar 

  • Neckers LM (1989): Antisense oligonucleotides as a tool for studying cell regulation: mechanism of uptake and application to the study of oncogene function. In: Oligodeoxynucleotides: Antisense inhibitors of gene expression, Cohen J, ed. London: Macmillan Press

    Google Scholar 

  • Pardridge WM, Boado RJ (1991): Enhanced cellular uptake of biotinylated antisense oligonucleotide or peptide mediated by avidin, a cationic protein. FEBS Letters 288: 30–32

    Article  PubMed  CAS  Google Scholar 

  • Pastan I, Willingham M, Anderson W, Gallo M (1977): Localization of serum-derived a2 macroglobulin in cultured cells and decrease after Moloney sarcoma virus transformation. Cell 609–617

    Google Scholar 

  • Pastan IH, Willingham MC (1981): Journey to the center of the cell: Role of the receptosome. Science 214: 504–509

    Article  PubMed  CAS  Google Scholar 

  • Renneisen K, Leserman L, Matthes E, Schroder HC, Muller WEG (1990): Inhibition of expression of human immunodefiency virus-1 in vitro by antibody-targeted liposomes containing antisense RNA to the env region. J Biol Chem 265: 16337–16342

    PubMed  CAS  Google Scholar 

  • Ryser HJ-P, Shen W-C (1978): Conjugation of methotrexate to poly(Llysine) increases drug transport and overcomes drug resistance in cultured cells. Proc Natl Acad Sci USA 75: 3867–3870

    Article  PubMed  CAS  Google Scholar 

  • Sburlati AR, Manrow RE, Berger SL (1991): Prothymosin a antisense oligomers inhibit myeloma cell division. Proc Natl Acad Sci USA 88: 253–257

    Article  PubMed  CAS  Google Scholar 

  • Shea RG, Marsters JC, Bischofberger N (1990): Synthesis, hybridization properties and antiviral activity of lipid-oligodeoxynucleotide conjugates. Nucl Acid Res 18: 3777–3783

    Article  CAS  Google Scholar 

  • Shoji Y, Akhtar S, Periasamy A, Herman B, Juliano RL (1991): Mechanism of cellular uptake of modified oligodeoynucleotides containing methylphosphonate linkages. Nucl Acid Res 19: 5543–5550

    Article  CAS  Google Scholar 

  • Stein CA, Mori K, Loke SL, Subasinghe K, Cohen JS, Neckers LM (1988): Phosphorothioate and normal oligodeoxyribonucleotides with 5′-linked acridine: characterization and preliminary kinetics of cellular uptake. Gene 72: 333–341

    Article  PubMed  CAS  Google Scholar 

  • Stevenson M, Iversen PL (1989): Inhibition of human immunodeficiency virus type l-mediated cytopathic effects by poly(L-lysine)-conjugated synthetic antisense oligodeoxyribonucleotides. J Gen Virol 70: 2673

    Article  PubMed  CAS  Google Scholar 

  • van Renswoude J, Bridges KR, Harford JB, Klausner RD (1982): Receptormediated endocytosis of transferrin and the uptake of Fe in K562 cells: identification of a nonlysosomal acidic compartment. Proc Natl Acad Sci USA 79: 6186–6190

    Article  PubMed  Google Scholar 

  • Willingham MC, Hanover JA, Dickson RB, Pastan I (1984): Morphologic characterization of the human pathway of transferrin endocytosis and recycling in human KB cells. Proc Natl Acad Sci USA 81: 175–179

    Article  PubMed  CAS  Google Scholar 

  • Willingham MC, Pastan I (1980): The receptosome: an intermediate organelle of receptor-mediated endocytosis in cultured fibroblasts. Cell 21: 67–77

    Article  PubMed  CAS  Google Scholar 

  • Yakubov LA, Deeva EA, Zarytova VF, Ivanova EM, Ryte AS, Yurchenko VL, Vlassov VV (1989): Mechanism of oligonucleotide uptake by cells: Involvement of specific receptors? Proc Natl Acad Sci USA 86: 6454–6458

    Article  PubMed  CAS  Google Scholar 

  • Zamecnik PC, Stephenson ML (1978): Inhibition of Rous sarcoma virus replication and cell transformation by a specific oligodeoxynucleotide. Proc Natl Acad Sci USA 75: 280–284

    Article  PubMed  CAS  Google Scholar 

  • Zhang XH, Loke SL, Neckers L (1990): unpublished observations

    Google Scholar 

Download references

Authors
  1. Leonard M. Neckers
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Department of Pediatrics, University of Wisconsin Medical School, Waisman Center, Room 607 1500 Highland Avenue, Madison, WI, 53705-2208, USA

    Jon A. Wolff

  2. Department of Medical Genetics, University of Wisconsin Medical School, Waisman Center, Room 607 1500 Highland Avenue, Madison, WI, 53705-2208, USA

    Jon A. Wolff

  3. Department of Neurology, University of Wisconsin Medical School, Waisman Center, Room 607 1500 Highland Avenue, Madison, WI, 53705-2208, USA

    Jon A. Wolff

Rights and permissions

Reprints and permissions

Copyright information

© 1994 Birkhäuser Boston

About this chapter

Cite this chapter

Neckers, L.M. (1994). Cellular Internalization of Oligodeoxynucleotides. In: Wolff, J.A. (eds) Gene Therapeutics. Birkhäuser Boston. https://doi.org/10.1007/978-1-4684-6822-9_10

Download citation

  • DOI: https://doi.org/10.1007/978-1-4684-6822-9_10

  • Publisher Name: Birkhäuser Boston

  • Print ISBN: 978-1-4684-6824-3

  • Online ISBN: 978-1-4684-6822-9

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics

Search

Navigation