Stratégie du leurre moléculaire en thérapie génique : ciblage de facteurs de transcription par un minicercle d'ADN multisites et étude de l'interaction entre l'ADN platiné et NF-κB

Abstract : In gene therapy, decoy strategy consists in using short nucleic acid containing a sequence specificallyrecognized by a transcription factor in order to trap it and in turn to inhibit the expression of genesunder its control. The therapeutic efficacy of this strategy is related to the biostability of the nucleicacid and the possibility to target several transcription factors that cooperate to induce cellderegulations responsible in human diseases. We have developed a technology to product in vitroDNA minicircle less than 250 base pairs which can interact with several proteins, chemicallyfunctionalizable and resistant to exonucleases. A dual targeting of the transcription factor NF-κB by aminicircle allows inhibition in cellulo of a NF-κB-dependent transcription reporter system consistentwith a decoy activity induced by the minicircle.Cisplatin is an anticancer drug whose pharmacological target is DNA. In several cancer cell lines, thedecrease of transcriptional activity of NF-κB has been linked to an increase in the sensitivity of cells tocisplatin. We studied in cellulo whether the mechanism of inhibition of NF-κB by cisplatin could dependon the platination of its consensus sequence using a novel application of the decoy strategy. We showthat cisplatin adducts, unlike those of transplatin (inactive isomer), decreases recognition of NF-κB toits consensus sequence in vitro and in cellulo.Decoy strategy with minicircle DNA will be developed to make the proof of concept of this approach atthe cellular level and further to demonstrate its biological effectiveness.
Document type :
Theses
Complete list of metadatas

https://tel.archives-ouvertes.fr/tel-01124106
Contributor : Abes Star <>
Submitted on : Friday, March 6, 2015 - 4:15:21 AM
Last modification on : Wednesday, June 27, 2018 - 8:24:02 AM
Long-term archiving on : Sunday, June 7, 2015 - 12:05:23 PM

File

2012ORLE2069.pdf
Version validated by the jury (STAR)

Identifiers

  • HAL Id : tel-01124106, version 1

Collections

Citation

Thomas Thibault. Stratégie du leurre moléculaire en thérapie génique : ciblage de facteurs de transcription par un minicercle d'ADN multisites et étude de l'interaction entre l'ADN platiné et NF-κB. Sciences agricoles. Université d'Orléans, 2012. Français. ⟨NNT : 2012ORLE2069⟩. ⟨tel-01124106⟩

Share

Metrics

Record views

440

Files downloads

823