利用伴侣蛋白@DNA 探针在细胞核和核仁中对人嘌呤/嘧啶内切核酸酶 1 进行活细胞成像。

Live-cell imaging of human apurinic/apyrimidinic endonuclease 1 in the nucleus and nucleolus using a chaperone@DNA probe.

机构信息

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

出版信息

Nucleic Acids Res. 2024 May 8;52(8):e41. doi: 10.1093/nar/gkae202.

DOI:10.1093/nar/gkae202

PMID:38554110

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11077052/

Abstract

Human apurinic/apyrimidinic endonuclease 1 (APE1) plays crucial roles in repairing DNA damage and regulating RNA in the nucleus. However, direct visualization of nuclear APE1 in live cells remains challenging. Here, we report a chaperone@DNA probe for live-cell imaging of APE1 in the nucleus and nucleolus in real time. The probe is based on an assembly of phenylboronic acid modified avidin and biotin-labeled DNA containing an abasic site (named PB-ACP), which cleverly protects DNA from being nonspecifically destroyed while enabling targeted delivery of the probe to the nucleus. The PB-ACP construct specifically detects APE1 due to the high binding affinity of APE1 for both avidin and the abasic site in DNA. It is easy to prepare, biocompatible and allowing for long-term observation of APE1 activity. This molecular tool offers a powerful means to investigate the behavior of APE1 in the nuclei of various types of live cells, particularly for the development of improved cancer therapies targeting this protein.

摘要

人类脱嘌呤/脱嘧啶核酸内切酶 1（APE1）在修复 DNA 损伤和调节细胞核内 RNA 方面发挥着关键作用。然而，直接在活细胞中可视化核 APE1 仍然具有挑战性。在这里，我们报告了一种伴侣@DNA 探针，用于实时在活细胞中对核内和核仁中的 APE1 进行成像。该探针基于苯硼酸修饰的亲和素和含有无碱基位点的生物素标记 DNA 的组装（命名为 PB-ACP），该探针巧妙地保护 DNA 免受非特异性破坏，同时能够将探针靶向递送至细胞核。由于 APE1 对亲和素和 DNA 中的无碱基位点具有高结合亲和力，因此 PB-ACP 构建体特异性地检测 APE1。它易于制备、生物相容性好，并允许对 APE1 活性进行长期观察。这种分子工具为研究各种类型活细胞中 APE1 的行为提供了一种强大的手段，特别是对于开发针对该蛋白的改进癌症治疗方法。