Based On Application Of Metal-organic Framework Nanoparticles And DNA Logic Gates In Living Cells

Cells,as the basic structure and functional unit of organisms,many specific biomarkers in the biological environment are closely relevant to the occurrence,differentiation and development of diseases,and are essential for the regulation of cell physiological functions.Exploring the changes in the expression levels of specific cancer biomolecules in living cells is of great guiding significance for the research on the prognosis and diagnosis of cancer in biomedicine.The diverse composition of cells makes the biological environment very complex.A disease is usually associated with one or more biomarkers or shares one biomarker with other diseases.Therefore,the application of a single biomarker detection method in the living cell for disease diagnosis is risky.DNA biological logic gates depend on the specificity and programmability of DNA to construct multiple types of logic gates.Therefore,constructing a DNA logic gate with a set of biomarkers as logic input to analyze changes the concentration of biomarkers in living cells can improve the accuracy of cancer cell identification.This paper used biomineralized metal-organic framework material as a carrier to design a series of logic gate systems that can work in living cells to achieve cell identification.The specific research content is as follows:In the second chapter,The polymerase-driven primer exchange reaction(PER)-DNAzyme cascade logic,system was developed to respond to two mi RNAs simultaneously in vitro.The DNA logic gate is composed of mi RNAs as input,primer exchange reaction as sensing module,Mg²⁺DNAzyme as processing module,and fluorescence signal as output.Three logic gates of AND,OR,and NOT are constructed.mi RNA-21 as the logic input,In the presence of polymerase,the primer exchange reaction autonomously generates logic components to trigger the downstream DNAzyme circuit to realize the response of mi RNAs in vitro.The polymerase-driven DNA biocomputing platform provides an innovative response mode,which further improves the programmability and accuracy of logic operations.Reprogramming the PER catalytic hairpin can be used to respond to other biomolecules.The introduction of polymerase makes the design expandable,but the implementation of this logic gate in living cells needs further study.In the third chapter,In order to realize that the biolog ical logic platform designed in Chapter 2 can work in living cells,we report for the first time that the metal organic framework nanomaterials through biomineralization serve as a carrier for co-delivering polymerase and DNA logic components into living c ells to initiate polymerase-driven PER-DNAzyme biocomputing platform(AND gate and OR gate)working in living cells to achieve reliable cell identification by detecting diverse endogenous mi RNAs.The results show that the bispecific DNA biologic logic platform built by us has programmability and accuracy,makes cell identification more reliable,meets the needs of biomedicine and its application in clinical diagnosis,and provides accurate diagnosis and treatment of diseases.The biomedical research of DNA logic gates in complex cell systems provides new tools and ideas.