Targeting the human androgen receptor gene with platinated triplex-forming oligonucleotides

Biochemistry
Mindy K GrahamP S Miller

Abstract

Platinum-derivatized homopyrimidine triplex-forming oligonucleotides (Pt-TFOs) consisting of 2'-O-methyl-5-methyluridine, 2'-O-methyl-5-methylcytidine, and a single 3'-N7-trans-chlorodiammine platinum(II)-2'-deoxyguanosine were designed to cross-link to the transcribed strand at four different sequences in the human androgen receptor (AR) gene. Fluorescence microscopy showed that a fluorescein-tagged Pt-TFO localizes in both the cytoplasm and nucleus when it is transfected into LAPC-4 cells, a human prostate cancer cell line, using Lipofectamine 2000. A capture assay employing streptavidin-coated magnetic beads followed by polymerase chain reaction (PCR) amplification was used to demonstrate that 5'-biotin-conjugated Pt-TFOs cross-link in vitro to their four designated AR gene targets in genomic DNA extracted from LAPC-4 cells. Similarly, the capture assay was used to examine cross-linking between the 5'-biotin-conjugated Pt-TFOs and the AR gene in LAPC-4 cells in culture. Three of the four Pt-TFOs cross-linked to their designated target, suggesting that different regions of the AR gene are not uniformly accessible to Pt-TFO cross-linking. LAPC-4 cells were transfected with fluorescein-tagged Pt-TFO or a control oligonucleotide...Continue Reading

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Citations

Apr 27, 2016·Chemistry : a European Journal·Dattatraya UkaleTuomas Lönnberg
Mar 27, 2018·Chembiochem : a European Journal of Chemical Biology·Dattatraya Uttam Ukale, Tuomas Lönnberg
Dec 12, 2017·Chemistry : a European Journal·Sajal Kumar Maity, Tuomas Lönnberg
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Mar 19, 2019·Current Protocols in Nucleic Acid Chemistry·Yosuke TaniguchiShigeki Sasaki
Aug 21, 2020·Chemical Communications : Chem Comm·Lei WangYosuke Taniguchi

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