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| Home > Publications database > Induction of specific chromosomal rearrangements by targeting sensitive genomic loci usingI-125-labeled Triplex-forming oligonucleotides |
| Home > Publications database > Induction of specific chromosomal rearrangements by targeting sensitive genomic loci usingI-125-labeled Triplex-forming oligonucleotides |
| Poster (After Call) | FZJ-2019-04706 |
; ;
2019
Please use a persistent id in citations: http://hdl.handle.net/2128/22815
Abstract: Triplex-Forming-oligonucleotides (TFO) bind DNA in a sequence-specific manner and possess therapeutic potential e.g. as a carrier-molecule for Auger-Electron-Emitter (AEE) targeting specific DNA sequences in tumor cells. We established a method for the labeling of TFO with the AEE Iodine-125 (125I) and analyzed the influence of 125I-labeled TFO in SCL-II cells on gene expression, translocation frequency and protein expression of the human BCL2 gene. TFO-BCL2 binds to the BCL2 gene upstream of the 3´-end. TFO labeling with 125I was performed using the primer-extension-method. SCL-II cells were transfected with TFO via electroporation and subsequently stored at -150°C for decay accumulation. SCL-II cells transfected with 125I-labeled multi-binding TFO or non-labeled TFO-BCL2 served as controls. Monitoring of BCL2 translocations was done with Fluorescence-In-Situ-Hybridization (FISH). The utilized FISH-probes were designed to detect a characteristic t(14;18) translocation of the BCL2 gene, leading to an overexpression of BCL-2. Gene expression levels were measured via qRT-PCR. Verification of gene expression on the protein level was analyzed by Western blotting. The relative gene expression of BCl-2 in 125I-TFO-BCL2 transfected cells showed a significant up-regulation and analysis of the BCL2 t(14;18) translocation frequency revealed a significant ~ 1.8-fold increase. This increase was not reflected on the protein level.We conclude that 125I decays within the BCL2 gene facilitate the t(14;18) chromosomal translocation and that the increased translocation frequency contributes to the observed enhanced BCL-2 gene expression. Funded by Bundesministerium für Bildung und Forschung (BMBF), Grant 02NUK005A and 02NUK043A
Keyword(s): Biology (2nd)
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