Orenstein, Alexander and Berlyoung, April and Rastede, Elizabeth and Pham, Ha and Fouquerel, Elise and Murphy, Connor and Leibowitz, Brian and Yu, Jian and Srivastava, Tumul and Armitage, Bruce and Opresko, Patricia
(2017)
γPNA FRET Pair Miniprobes for Quantitative Fluorescent In Situ Hybridization to Telomeric DNA in Cells and Tissue.
Molecules, 22 (12).
ISSN 1420-3049
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.
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| Item Type: |
Article
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| Status: |
Published |
| Creators/Authors: |
| Creators | Email | Pitt Username | ORCID  |
|---|
| Orenstein, Alexander | | | | | Berlyoung, April | | | | | Rastede, Elizabeth | | | | | Pham, Ha | | | | | Fouquerel, Elise | | | | | Murphy, Connor | | | | | Leibowitz, Brian | bjl40@pitt.edu | bjl40 | | | Yu, Jian | jiy3@pitt.edu | jiy3 | | | Srivastava, Tumul | | | | | Armitage, Bruce | | | | | Opresko, Patricia | plo4@pitt.edu | plo4 | |
|
| Date: |
2 December 2017 |
| Date Type: |
Publication |
| Journal or Publication Title: |
Molecules |
| Volume: |
22 |
| Number: |
12 |
| Publisher: |
MDPI AG |
| DOI or Unique Handle: |
10.3390/molecules22122117 |
| Schools and Programs: |
School of Public Health > Environmental and Occupational Health |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
telomere, γPNA, hybridization probe, fluorescent imaging |
| ISSN: |
1420-3049 |
| Official URL: |
http://dx.doi.org/10.3390/molecules22122117 |
| Funders: |
NIH grants, David Scaife Family Charitable Foundation |
| Article Type: |
Research Article |
| Date Deposited: |
13 Jan 2021 18:56 |
| Last Modified: |
13 Jan 2021 18:56 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/40146 |
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