Nucleic Acids Research, Vol 24, Issue 2 295-302, Copyright © 1996 by Oxford University Press
F Svinarchuk, A Debin, JR Bertrand and C Malvy
In our previous work we have shown that the oligonucleotide 5'-
GGGGAGGGGGAGG-3' gives a very stable and specific triplex with the promoter
of the murine c-pim-1 proto-oncogene in vitro[Svinarchuk, F., Bertrand,
J.-R. and Malvy, C.(1994)Nucleic Acids Res., 22, 3742-3747]. In the present
work, we have tested triplex formation with some derivatives of this
oligonucleotide which are designed to be degradation-resistant inside the
cells, and we show that phosphorothioate and the oligonucleotide with a 3'
terminal amino group are still able to form triplexes. Moreover these
oligonucleotides, like the 13mer oligonucleotide of similar composition
[Svinarchuk, F., Paoletti, J., and Malvy, C. (1995) J. Biol. Chem., 270,
14068-14071], are able to stabilize the targeted duplex. In vivo DMS
footprint analysis after electroporation of the pre-formed triplex into the
cell have shown the presence of the triple helix inside the cells. This
triplex structure partially blocks c-pim-1 promotor activity as shown by
transient assay with a c-pim-1 promoter-luciferase gene construct. To our
knowledge these data are the first direct evidence that conditions inside
cells are favorable for triplex stability with non- modified
oligonucleotides. However we were unable to show triplex formation inside
living cells using various methods of oligonucleotide delivery. We suppose
that this may be due to the oligonucleotide being sequestered by cellular
processes or proteins. Further work is needed to find oligonucleotide
derivatives and ways of their delivery to overcome the problem of triplex
formation inside the cells.
ARTICLES
Investigation of the intracellular stability and formation of a triple helix formed with a short purine oligonucleotide targeted to the murine c-pim-1 proto-oncogene promotor
Laboratoire de Biochimie-Enzymologie, CNRS URA 147, Institute Gustave Roussy, Villejuif, France.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  C. Shen, A. Buck, G. Mehrke, B. Polat, H.-J. Gross, M. Bachem, and S. Reske Triplex forming oligonucleotide targeted to 3'UTR downregulates the expression of the bcl-2 proto-oncogene in HeLa cells Nucleic Acids Res., February 1, 2001; 29(3): 622 - 628. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z. Intody, B. D. Perkins, J. H. Wilson, and T. G. Wensel Blocking transcription of the human rhodopsin gene by triplex-mediated DNA photocrosslinking Nucleic Acids Res., November 1, 2000; 28(21): 4283 - 4290. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Bailey and D. L. Weeks Understanding oligonucleotide-mediated inhibition of gene expression in Xenopus laevis oocytes Nucleic Acids Res., March 1, 2000; 28(5): 1154 - 1161. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. Giovannangeli, S. Diviacco, V. Labrousse, S. Gryaznov, P. Charneau, and C. Helene Accessibility of nuclear DNA to triplex-forming oligonucleotides: The integrated HIV-1 provirus as a target PNAS, January 7, 1997; 94(1): 79 - 84. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 F.-L. M. Lin, A. Majumdar, L. C. Klotz, A. P. Reszka, S. Neidle, and M. M. Seidman Stability of DNA Triplexes on Shuttle Vector Plasmids in the Replication Pool in Mammalian Cells J. Biol. Chem., December 8, 2000; 275(50): 39117 - 39124. [Abstract] [Full Text] [PDF]
|
|