Fluorescence lifetime imaging for studying DNA compaction and gene activities

Svitlana M. Levchenko; Artem Pliss; Xiao Peng; Paras N. Prasad; Junle Qu

doi:10.1038/s41377-021-00664-w

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Fluorescence lifetime imaging for studying DNA compaction and gene activities

Articles

- Fluorescence lifetime imaging for studying DNA compaction and gene activities
- Fluorescence lifetime imaging for studying DNA compaction and gene activities
- LSA 2021年10卷第12期页码：2349-2359
- Affiliations：
  
  1.Key Laboratory of Optoelectronic Devices and Systems of Ministry of Education and Guangdong Province, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen, Guangdong 518060, China
  2.Department of Cell Biophysics, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland
  3.Institute for Lasers, Photonics and Biophotonics, University at Buffalo, State University of New York, Buffalo, NY 14260-3000, USA
- Author bio：
  
  Paras N. Prasad (pnprasad@buffalo.edu)
  Junle Qu (jlqu@szu.edu.cn)
- Funds：
- DOI：10.1038/s41377-021-00664-w
  中图分类号：
- 纸质出版日期：2021-12-31，
  
  网络出版日期：2021-11-02，
  
  收稿日期：2021-03-25，
  
  修回日期：2021-10-04，
  
  录用日期：2021-10-18
- Accepted：
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Fluorescence lifetime imaging for studying DNA compaction and gene activities[J]. LSA, 2021,10(12):2349-2359.

Levchenko, S. M. et al. Fluorescence lifetime imaging for studying DNA compaction and gene activities. Light: Science & Applications, 10, 2349-2359 (2021).
Fluorescence lifetime imaging for studying DNA compaction and gene activities[J]. LSA, 2021,10(12):2349-2359. DOI： 10.1038/s41377-021-00664-w.

Levchenko, S. M. et al. Fluorescence lifetime imaging for studying DNA compaction and gene activities. Light: Science & Applications, 10, 2349-2359 (2021). DOI： 10.1038/s41377-021-00664-w.

摘要

Abstract

Optical imaging is a most useful and widespread technique for the investigation of the structure and function of the cellular genomes. However

an analysis of immensely convoluted and irregularly compacted DNA polymer is highly challenging even by modern super-resolution microscopy approaches. Here we propose fluorescence lifetime imaging (FLIM) for the advancement of studies of genomic structure including DNA compaction

replication as well as monitoring of gene expression. The proposed FLIM assay employs two independent mechanisms for DNA compaction sensing. One mechanism relies on the inverse quadratic relation between the fluorescence lifetimes of fluorescence probes incorporated into DNA and their local refractive index

variable due to DNA compaction density. Another mechanism is based on the Förster resonance energy transfer (FRET) process between the donor and the acceptor fluorophores

both incorporated into DNA. Both these proposed mechanisms were validated in cultured cells. The obtained data unravel a significant difference in compaction of the gene-rich and gene-poor pools of genomic DNA. We show that the gene-rich DNA is loosely compacted compared to the dense DNA domains devoid of active genes.

关键词

Keywords

references

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