Telomere Visualization in Tissue Sections using Pyrrole–Imidazole Polyamide Probes
نویسندگان
چکیده
Pyrrole-Imidazole (PI) polyamides bind to specific DNA sequences in the minor groove with high affinity. Specific DNA labeling by PI polyamides does not require DNA denaturation with harsh treatments of heat and formamide and has the advantages of rapid and less disruptive processing. Previously, we developed tandem hairpin PI polyamide probes (TH59 series), which label telomeres in cultured cell lines more efficiently than conventional methods, such as fluorescence in situ hybridization (FISH). Here, we demonstrate that a TH59 derivative, HPTH59-b, along with immunostaining for specifying cell types in the tissues, visualizes telomeres in mouse and human tissue sections. Quantitative measurements of telomere length with single-cell resolution suggested shorter telomeres in the proliferating cell fractions of tumor than in non-tumor tissues. Thus, PI polyamides are a promising alternative for telomere labeling in clinical research, as well as in cell biology.
منابع مشابه
Tandem trimer pyrrole–imidazole polyamide probes targeting 18 base pairs in human telomere sequences† †Electronic supplementary information (ESI) available: Materials and methods, sequences of ODN-1 to 5, mass spectra and analytical HPLC profiles of the compounds TH59, TT59, TAMRA TH59 and TAMRA TT59, the UV-visible absorption and fluorescence spectra of probes TAMRA TH59 and TAMRA TT59, SPR sensorgrams of TH59 and TT59, additional images of telomeres. See DOI: 10.1039/c4sc03755c Click here for additional data file.
متن کامل
Tandem trimer pyrrole-imidazole polyamide probes targeting 18 base pairs in human telomere sequences.
The binding of molecules to specific DNA sequences is important for imaging genome DNA and for studying gene expression. Increasing the number of base pairs targeted by these molecules would provide greater specificity. N-Methylpyrrole-N-methylimidazole (Py-Im) polyamides are one type of such molecules and can bind to the minor groove of DNA in a sequence-specific manner without causing denatur...
متن کاملDevelopment of Pyrrole-Imidazole (PI) Polyamides as
Disclosed is an inhibitor of the expression of CTGF gene, which comprises a pyrrole-imidazole polyamide. The pyrrole-imidazole polyamide has an N-methylpyrrole unit (hereinafter, referred to as “Py”), an N-methylimidazole unit (hereinafter, referred to as “Im”) and a ϝ-aminobutanoic acid unit. In the pyrrole-imidazole polyamide, the region of the ϝ-aminobutanoic acid unit is folded to form an U...
متن کاملIn vivo imaging of pyrrole-imidazole polyamides with positron emission tomography.
The biodistribution profiles in mice of two pyrrole-imidazole polyamides were determined by PET. Pyrrole-imidazole polyamides are a class of small molecules that can be programmed to bind a broad repertoire of DNA sequences, disrupt transcription factor-DNA interfaces, and modulate gene expression pathways in cell culture experiments. The (18)F-radiolabeled polyamides were prepared by oxime lig...
متن کاملA sequence-specific DNA binding small molecule triggers the release of immunogenic signals and phagocytosis in a model of B-cell lymphoma.
Means to cause an immunogenic cell death could lead to significant insight into how cancer escapes immune control. In this study, we screened a library of five pyrrole-imidazole polyamides coding for different DNA sequences in a model of B-cell lymphoma for the upregulation of surface calreticulin, a pro-phagocytosis signal implicated in immunogenic cell death. We found that hairpin polyamide 1...
متن کامل