Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy

Raymond Bergan, Yvette Connell, Brigid Fahmy, Leonard Neckers

Research output: Contribution to journalArticle

100 Citations (Scopus)

Abstract

Obtaining high transfection efficiencies and achieving appropriate intracellular concentrations and localization are two of the most important barriers to the implementation of gene targeted therapy. The efficiency of endogenous uptake of oligodeoxynucleotides (ODNs) varies from cell type to cell type and may be a limiting factor of antisense efficacy. The use of electroporation to obtain high intracellular concentrations of a synthetic ODN in essentially 100% of viable cells is described. It is also shown that the transfected ODNs initially localize to the nucleus and remain there for at least 48 hours. The cellular trafficking of electroporated ODNs is shown to be an energy dependent process. Targeting of the c-myc proto-oncogene of U937 cells by electroporation of phosphorothioate-modified ODNs results in rapid and specific suppression of this gene at ODN concentrations much lower than would otherwise be required. This technique appears to be applicable to a variety of cell types and may represent a powerful new investigative tool as well as a promising approach to the ex vivo treatment of hematologic disorders.

Original languageEnglish (US)
Pages (from-to)3567-3573
Number of pages7
JournalNucleic Acids Research
Volume21
Issue number15
StatePublished - Jul 25 1993
Externally publishedYes

Fingerprint

Electroporation
Oligodeoxyribonucleotides
Efficacy
Cell
Gene Therapy
Genes
U937 Cells
myc Genes
Nucleus
High Efficiency
Disorder
Genetic Therapy
Limiting
Transfection
Vary
Gene
Dependent
Energy
Factors
Localization

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Bergan, R., Connell, Y., Fahmy, B., & Neckers, L. (1993). Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy. Nucleic Acids Research, 21(15), 3567-3573.

Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy. / Bergan, Raymond; Connell, Yvette; Fahmy, Brigid; Neckers, Leonard.

In: Nucleic Acids Research, Vol. 21, No. 15, 25.07.1993, p. 3567-3573.

Research output: Contribution to journalArticle

Bergan, R, Connell, Y, Fahmy, B & Neckers, L 1993, 'Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy', Nucleic Acids Research, vol. 21, no. 15, pp. 3567-3573.
Bergan, Raymond ; Connell, Yvette ; Fahmy, Brigid ; Neckers, Leonard. / Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy. In: Nucleic Acids Research. 1993 ; Vol. 21, No. 15. pp. 3567-3573.
@article{f0c25239886248efa8f51c9237e57646,
title = "Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy",
abstract = "Obtaining high transfection efficiencies and achieving appropriate intracellular concentrations and localization are two of the most important barriers to the implementation of gene targeted therapy. The efficiency of endogenous uptake of oligodeoxynucleotides (ODNs) varies from cell type to cell type and may be a limiting factor of antisense efficacy. The use of electroporation to obtain high intracellular concentrations of a synthetic ODN in essentially 100{\%} of viable cells is described. It is also shown that the transfected ODNs initially localize to the nucleus and remain there for at least 48 hours. The cellular trafficking of electroporated ODNs is shown to be an energy dependent process. Targeting of the c-myc proto-oncogene of U937 cells by electroporation of phosphorothioate-modified ODNs results in rapid and specific suppression of this gene at ODN concentrations much lower than would otherwise be required. This technique appears to be applicable to a variety of cell types and may represent a powerful new investigative tool as well as a promising approach to the ex vivo treatment of hematologic disorders.",
author = "Raymond Bergan and Yvette Connell and Brigid Fahmy and Leonard Neckers",
year = "1993",
month = "7",
day = "25",
language = "English (US)",
volume = "21",
pages = "3567--3573",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "15",

}

TY - JOUR

T1 - Electroporation enhances c-myc antisense oligodeoxynucleotide efficacy

AU - Bergan, Raymond

AU - Connell, Yvette

AU - Fahmy, Brigid

AU - Neckers, Leonard

PY - 1993/7/25

Y1 - 1993/7/25

N2 - Obtaining high transfection efficiencies and achieving appropriate intracellular concentrations and localization are two of the most important barriers to the implementation of gene targeted therapy. The efficiency of endogenous uptake of oligodeoxynucleotides (ODNs) varies from cell type to cell type and may be a limiting factor of antisense efficacy. The use of electroporation to obtain high intracellular concentrations of a synthetic ODN in essentially 100% of viable cells is described. It is also shown that the transfected ODNs initially localize to the nucleus and remain there for at least 48 hours. The cellular trafficking of electroporated ODNs is shown to be an energy dependent process. Targeting of the c-myc proto-oncogene of U937 cells by electroporation of phosphorothioate-modified ODNs results in rapid and specific suppression of this gene at ODN concentrations much lower than would otherwise be required. This technique appears to be applicable to a variety of cell types and may represent a powerful new investigative tool as well as a promising approach to the ex vivo treatment of hematologic disorders.

AB - Obtaining high transfection efficiencies and achieving appropriate intracellular concentrations and localization are two of the most important barriers to the implementation of gene targeted therapy. The efficiency of endogenous uptake of oligodeoxynucleotides (ODNs) varies from cell type to cell type and may be a limiting factor of antisense efficacy. The use of electroporation to obtain high intracellular concentrations of a synthetic ODN in essentially 100% of viable cells is described. It is also shown that the transfected ODNs initially localize to the nucleus and remain there for at least 48 hours. The cellular trafficking of electroporated ODNs is shown to be an energy dependent process. Targeting of the c-myc proto-oncogene of U937 cells by electroporation of phosphorothioate-modified ODNs results in rapid and specific suppression of this gene at ODN concentrations much lower than would otherwise be required. This technique appears to be applicable to a variety of cell types and may represent a powerful new investigative tool as well as a promising approach to the ex vivo treatment of hematologic disorders.

UR - http://www.scopus.com/inward/record.url?scp=0027242052&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027242052&partnerID=8YFLogxK

M3 - Article

VL - 21

SP - 3567

EP - 3573

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 15

ER -