研究目的
Investigating the effect of bond-cleavage type and position on the metabolic pathway of the EdU derivatives in living systems.
研究成果
The derivative linked by methylene at the N-3 position of EdU was not involved in metabolism without photodecaging, while the carbonate-linked derivative at the 3' oxygen atom could participate in metabolism in vivo without photodecaging. This research provides a reference for evaluating whether small molecules are involved in metabolism after photocaging modification.
研究不足
The stability and photodecaging efficiency of the photocaged EdU derivatives vary depending on the attachment site of the photocage group, which could affect their metabolic pathway in living systems.
1：Experimental Design and Method Selection:
Designed two photocaged 5-ethynyl-2′-deoxyuridine (EdU) derivatives to systematically investigate the effect of bond-cleavage type and position on the metabolic pathway.
2：Sample Selection and Data Sources:
Used HeLa cells for the experiments.
3：List of Experimental Equipment and Materials:
Included HPLC for analysis, UV irradiation at 365 nm for photodecaging, and fluorescent confocal microscopy for imaging.
4：Experimental Procedures and Operational Workflow:
Incubated photocage-EdU derivatives with cells, followed by UV irradiation, fixation, permeabilization, and staining with Alexa488 and DAPI for imaging.
5：Data Analysis Methods:
Analyzed photodecaging efficiency by HPLC, and genomic DNA modification by dot blot and mass spectrometry.
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