The Detection of Oligodeoxynucleotide Molecules Following Uptake into Mammalian Cells
- 1 January 1992
- journal article
- research article
- Published by Mary Ann Liebert Inc in Antisense Research and Development
- Vol. 2 (2) , 119-127
- https://doi.org/10.1089/ard.1992.2.119
Abstract
A mixed phosphodiester:phosphorothioate oligodeoxynucleotide was used in uptake studies with T15 mouse fibroblast cells. The presence of full-length unlabeled oligomers was identified in both cytoplasmic and nuclear extracts by a method involving gel electrophoresis and electroblotting followed by hybridization with a complementary radiolabeled probe. Detection did not depend on the presence of a label on the oligomer with the potential for its removal, often a problem in other studies. The fate of the oligonucleotides could then be followed with time for at least 3 days. This method of detection should be applicable to studies of nuclease resistance and uptake characteristics of newly developed oligonucleotide analogues.Keywords
This publication has 21 references indexed in Scilit:
- Evidence that a triplex-forming oligodeoxyribonucleotide binds to the c-myc promoter in HeLa cells, thereby reducing c-myc mRNA levels.Proceedings of the National Academy of Sciences, 1991
- Antisense inhibition of ras p21 expression that is sensitive to a point mutationBiochemistry, 1991
- Modified deoxyoligonucleotides stable to exonuclease degradation in serumNucleic Acids Research, 1991
- Mechanism of oligonucleotide uptake by cells: involvement of specific receptors?Proceedings of the National Academy of Sciences, 1989
- Synthesis of alkylating oligonucleotide derivatives containing cholesterol or phenazinium residues at their 3′‐terminus and their interaction with DNA within mammalian cellsFEBS Letters, 1989
- Regulation of viral expression of human immunodeficiency virus in vitro by an antisense phosphorothioate oligodeoxynucleotide against rev (art/trs) in chronically infected cells.Proceedings of the National Academy of Sciences, 1989
- Characterization of oligonucleotide transport into living cells.Proceedings of the National Academy of Sciences, 1989
- The inhibition of neunte outgrowth in PC12 cells by tubulin antisense oligodeoxyribonucleotidesGene, 1988
- Biochemical and biological effects of nonionic nucleic acid methylphosphonatesBiochemistry, 1981
- A rapid, sensitive, and versatile assay for protein using Coomassie brilliant blue G250Analytical Biochemistry, 1977