Affiliation:
1. State Key Laboratory of Organic‐Inorganic Composites Beijing Key Laboratory of Bioprocess Beijing Advanced Innovation Center for Soft Matter Science and Engineering College of Life Science and Technology Beijing University of Chemical Technology Beijing 100029 China
2. Sinopec Key Laboratory of Research and Application of Medical and Hygienic Materials Sinopec (Beijing) Research Institute of Chemical Industry Co., Ltd. Beijing 100013 China
3. State Key Laboratory of Pathogen and Biosecurity Academy of Military Medical Sciences Beijing 100071 China
Abstract
AbstractComplex temporal molecular signals play a pivotal role in the intricate biological pathways of living organisms, and cells exhibit the ability to transmit and receive information by intricately managing the temporal dynamics of their signaling molecules. Although biomimetic molecular networks are successfully engineered outside of cells, the capacity to precisely manipulate temporal behaviors remains limited. In this study, the catalysis activity of isothermal DNA polymerase (DNAP) through combined use of molecular dynamics simulation analysis and fluorescence assays is first characterized. DNAP‐driven delay in signal strand release ranged from 100 to 102 min, which is achieved through new strategies including the introduction of primer overhangs, utilization of inhibitory reagents, and alteration of DNA template lengths. The results provide a deeper insight into the underlying mechanisms of temporal control DNAP‐mediated primer extension and DNA strand displacement reactions. Then, the regulated DNAP catalysis reactions are applied in temporal modulation of downstream DNA‐involved reactions, the establishment of dynamic molecular signals, and the generation of barcodes for multiplexed detection of target genes. The utility of DNAP‐based signal delay as a dynamic DNA nanotechnology extends beyond theoretical concepts and achieves practical applications in the fields of cell‐free synthetic biology and bionic sensing.
Funder
National Natural Science Foundation of China