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基于G-Triplex辅助滚动圆放大功能的超高分辨率DNA中局部特异的8-Oxo-7,8-dihydroguanine的识别和定量

Jia-Hui Dong¹, Run-Hong Zhang¹, Ling-Li Zhao²

¹Beijing National Laboratory for Molecular Sciences (BNLMS), MOE Key Laboratory of Bioorganic Chemistry and Molecular Engineering, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.

Analytical chemistry

|December 27, 2023

概括

这项研究开发了一种敏感的方法来检测8-oxo-7,8-dihydroguanine (OG),这是一个重要的DNA损伤生物标志物. 该技术精确地量化了特定DNA位置的OG,即使在高水平的瓜干扰下也是如此.

科学领域:

生物化学生物化学
分子生物学分子生物学
遗传学是一个遗传学.

背景情况:

反应性氧物种 (ROS) 会导致与疾病,衰老和癌症有关的DNA损伤.
8-Oxo-7,8-dihydroguanine (OG) 是关键的瓜氧化产物和氧化应激和基因调节的生物标志物.
网站特定的OG量化对于理解其在转录激活和突变诱导中的双重作用至关重要.

研究的目的:

开发一种超灵敏的方法,用于定位特定量化8-oxo-7,8-dihydroguanine (OG).
克服在基因组DNA中区分低丰度OG与高丰度关氨酸 (G) 的挑战.
为了能够在氧化应激下监测基因组DNA中的OG,并识别具有功能意义的OG位点.

主要方法:

使用滚动圆放大 (RCA) 诱导G-三倍体形成.
用于信号检测的提奥夫拉T光增强剂.
纳入了DNA探针的酶性结合,以获得G背景信号的特异性和抑制.

主要成果:

实现了OG的超敏感检测,具有超低的检测极限 (0.18 amol).
在存在过量的G时,成功区分了0.05%的OG.
在氧化应激下,在基因组DNA中展示了特定于位点的OG的定性和定量监测.

结论:

开发的战略提供了一个高度敏感和特定的方法,用于特定位置的OG量化.
这种技术对于研究OG在DNA损伤和基因调节中的生物学作用非常有价值.
该方法有助于识别关键的OG部位,这些部位与疾病发病和细胞对氧化应激反应有关.

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