posted on 2024-01-03, 21:29authored byChen Wang, Zuoling Zhang, Yufei Qiu, Yaofei Bao, Qinxin Song, Bingjie Zou
In situ monitoring of the actions of correlated
enzymes in living cells is crucial for expanding our understanding
of disease progression and evaluating drug efficacy. However, due
to the diverse functions of different enzymes, currently available
methods for comprehensive analysis of these events are limited. Here,
we present an in situ track-generated DNA walker
for AND-gate logic imaging of telomerase (TE) and flap endonuclease
1 (FEN1) activities in live cells. TE is in charge of generating the
tracks for the walking strands by extending the TE primer on a gold
nanoparticle, while FEN1 is responsible for recognizing the overlapping
structure formed by the walking strands and the tracks and then cleaving
the fluorescent reporter to produce signals. By utilizing the DNA
walker, we successfully determined the expression levels and activities
of TE and FEN1 in various cancer cell lines, offering promising prospects
for screening inhibitors and investigating the biomolecular mechanisms
of diseases.