A novel DNA double-strand breaks biosensor based on fluorescence resonance energy transfer.

IF 11.3 1区 医学 Q1 Medicine Biomaterials Research Pub Date : 2023-02-17 DOI:10.1186/s40824-023-00354-1
Jung-Soo Suh, Tae-Jin Kim
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Abstract

Revealing the spatiotemporal behavior of DNA double-strand breaks (DSBs) is crucial for understanding the processes of DNA damage and repair. Traditionally, γH2AX and DNA damage response (DDR) factors have been used to detect DSBs using classical biochemical assays, such as antibody-based immunostaining. However, a reliable method to visualize and assess DSB activity real-time in living cells is yet to be established. Herein, we developed a novel DNA double-strand breaks biosensor (DSBS) based on fluorescence resonance energy transfer (FRET) by employing the H2AX and BRCT1 domains. By applying FRET imaging with DSBS, we show that DSBS specifically reacts to drug- or ionizing radiation (IR)-induced γH2AX activity, allowing for the quantification of DSB events at high spatiotemporal resolutions. Taken together, we provide a new experimental tool to evaluate the spatiotemporal dynamics of DNA double-strand breaks. Ultimately, our biosensor can be useful for elucidating the molecular mechanisms underlying DNA damage and repair processes.

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基于荧光共振能量转移的新型DNA双链断裂生物传感器。
揭示DNA双链断裂(DSBs)的时空行为对于理解DNA损伤和修复过程至关重要。传统上,使用γ - h2ax和DNA损伤反应(DDR)因子使用传统的生化分析,如基于抗体的免疫染色来检测dsb。然而,一种可靠的方法来可视化和实时评估DSB活性的活细胞尚未建立。在此,我们利用H2AX和BRCT1结构域开发了一种基于荧光共振能量转移(FRET)的DNA双链断裂生物传感器(DSBS)。通过应用DSBS的FRET成像,我们发现DSBS对药物或电离辐射(IR)诱导的γ - h2ax活性有特异性反应,从而可以在高时空分辨率下量化DSB事件。总之,我们提供了一个新的实验工具来评估DNA双链断裂的时空动力学。最终,我们的生物传感器可以用于阐明DNA损伤和修复过程的分子机制。
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来源期刊
Biomaterials Research
Biomaterials Research Medicine-Medicine (miscellaneous)
CiteScore
10.20
自引率
3.50%
发文量
63
审稿时长
30 days
期刊介绍: Biomaterials Research, the official journal of the Korean Society for Biomaterials, is an open-access interdisciplinary publication that focuses on all aspects of biomaterials research. The journal covers a wide range of topics including novel biomaterials, advanced techniques for biomaterial synthesis and fabrication, and their application in biomedical fields. Specific areas of interest include functional biomaterials, drug and gene delivery systems, tissue engineering, nanomedicine, nano/micro-biotechnology, bio-imaging, regenerative medicine, medical devices, 3D printing, and stem cell research. By exploring these research areas, Biomaterials Research aims to provide valuable insights and promote advancements in the biomaterials field.
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