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DNA binding fluorescent proteins as single-molecule probes

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Abstract

DNA binding fluorescent proteins are useful probes for a broad range of biological applications. Fluorescent protein (FP)-tagging allows DNA binding proteins expressed within a living cell to be directly visualised, in real-time, to study DNA binding patterns and dynamics. Moreover, FP-tagged DNA binding proteins (FP-DBP) have allowed the imaging of single proteins bound to large elongated DNA molecules with a fluorescence microscope. Although there are numerous DNA binding proteins, only a small portion of them have been exploited to construct FP-DBPs to study molecular motion in a cell or in vitro single-molecule visualisation. Therefore, it would be informative to review FP-DBP for further development. Here, we summarise the design of FP-DBPs and their brightness, photostability, pKa, maturation rate, and binding affinity (Kd) characteristics. Then, we review the applications of FP-DBP in cells to study chromosome dynamics, DNA replication, transcription factors, DNA damage, and repair. Finally, we focus on single DNA molecule visualisation using FP-DBP.

Graphical abstract: DNA binding fluorescent proteins as single-molecule probes

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Article information


Submitted
02 Feb 2020
Accepted
18 Apr 2020
First published
28 Apr 2020

Analyst, 2020, Advance Article
Article type
Critical Review

DNA binding fluorescent proteins as single-molecule probes

X. Jin, N. D. Hapsari, S. Lee and K. Jo, Analyst, 2020, Advance Article , DOI: 10.1039/D0AN00218F

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