Fluorescence imaging to understand the molecular mechanism of DNA damage-triggered cellular reprogramming in plants

Yosuke Tamada; Akihiro Imai; Nan Gu

doi:10.1117/12.2615404

27 October 2021 Fluorescence imaging to understand the molecular mechanism of DNA damage-triggered cellular reprogramming in plants

Yosuke Tamada, Akihiro Imai, Nan Gu

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Proceedings Volume 11925, Biomedical Imaging and Sensing Conference 2021; 1192506 (2021) https://doi.org/10.1117/12.2615404
Event: SPIE Technologies and Applications of Structured Light, 2021, Online, Japan

Abstract

We recently found that massive but transient DNA strand breaks, a kind of DNA damage, can trigger cellular reprogramming of differentiated cells to stem cells in the moss Physcomitrella patens (Physcomitrella). In Physcomitrella, DNA is massively damaged by the treatment with a DNA strand break-inducing reagent at high concentration. The DNA damage was repaired within 24 hours after the removal of the reagent. Then, the promoter activity of STEMIN1, encoding an integrator of the reprogramming signals, became active in a part of leaf cells, which eventually became stem cells. Levels of DNA strand breaks were quantified by performing comet assay, where the nuclei are subject to electrophoresis, stained with the fluorescent dye that binds to DNA, and observed with fluorescent microscope. The promoter activity of STEMIN1 was visualized using fluorescent protein and observed with fluorescent microscopy.

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Yosuke Tamada, Akihiro Imai, and Nan Gu "Fluorescence imaging to understand the molecular mechanism of DNA damage-triggered cellular reprogramming in plants", Proc. SPIE 11925, Biomedical Imaging and Sensing Conference 2021, 1192506 (27 October 2021); https://doi.org/10.1117/12.2615404

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KEYWORDS

Stem cells

Comets

Luminescence

Molecular mechanisms

Double sideband modulation

Head

Microscopes

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