Live-cell imaging of human apurinic/apyrimidinic endonuclease 1 in the nucleus and nucleolus using a chaperone@DNA probe.
Xiangjian CaoJinghui ZhengRuilan ZhangYing SunMeiping ZhaoPublished in: Nucleic acids research (2024)
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.
Keyphrases
- circulating tumor
- single molecule
- cell free
- living cells
- dna damage
- induced apoptosis
- endothelial cells
- nucleic acid
- high resolution
- quantum dots
- dna repair
- heavy metals
- cell cycle arrest
- oxidative stress
- risk assessment
- heat shock protein
- circulating tumor cells
- cell death
- endoplasmic reticulum stress
- signaling pathway
- small molecule
- cell proliferation
- aqueous solution