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Molecules 2017, 22(12), 2117; doi:10.3390/molecules22122117

γPNA FRET Pair Miniprobes for Quantitative Fluorescent In Situ Hybridization to Telomeric DNA in Cells and Tissue

1
Department of Environmental and Occupational Health, University of Pittsburgh Graduate School of Public Health, UPMC Hillman Cancer Center, Pittsburgh, PA 15213, USA
2
Department of Chemistry, Carnegie Mellon University, Pittsburgh, PA 15213, USA
3
Department of Pathology, University of Pittsburgh School of Medicine, UPMC Hillman Cancer Center, Pittsburgh, PA 15213, USA
4
Department of Radiation Oncology, University of Pittsburgh School of Medicine, UPMC Hillman Cancer Center, Pittsburgh, PA 15213, USA
5
PNA Innovations, Inc., 10-N Roessler Rd., Woburn, MA 01801, USA
6
Center for Nucleic Acids Science and Technology, Carnegie Mellon University, 4400 Fifth Avenue, Pittsburgh, PA 15213, USA
*
Authors to whom correspondence should be addressed.
Received: 27 October 2017 / Revised: 24 November 2017 / Accepted: 29 November 2017 / Published: 2 December 2017
(This article belongs to the Special Issue Molecular Properties and the Applications of Peptide Nucleic Acids)
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Abstract

Measurement of telomere length by fluorescent in situ hybridization is widely used for biomedical and epidemiological research, but there has been relatively little development of the technology in the 20 years since it was first reported. This report describes the use of dual gammaPNA (γPNA) probes that hybridize at alternating sites along a telomere and give rise to Förster resonance energy transfer (FRET) signals. Bright staining of telomeres is observed in nuclei, chromosome spreads and tissue samples. The use of FRET detection also allows for elimination of wash steps, normally required to remove unhybridized probes that would contribute to background signals. We found that these wash steps can diminish the signal intensity through the removal of bound, as well as unbound probes, so eliminating these steps not only accelerates the process but also enhances the quality of staining. Thus, γPNA FRET pairs allow for brighter and faster staining of telomeres in a wide range of research and clinical formats. View Full-Text
Keywords: telomere; γPNA; hybridization probe; fluorescent imaging telomere; γPNA; hybridization probe; fluorescent imaging
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Orenstein, A.; Berlyoung, A.S.; Rastede, E.E.; Pham, H.H.; Fouquerel, E.; Murphy, C.T.; Leibowitz, B.J.; Yu, J.; Srivastava, T.; Armitage, B.A.; Opresko, P.L. γPNA FRET Pair Miniprobes for Quantitative Fluorescent In Situ Hybridization to Telomeric DNA in Cells and Tissue. Molecules 2017, 22, 2117.

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