Research group of Professor Yang Dayong of Tianjin University: gene circuit compartmentalization on nano interface promotes cascade gene expression

In vivo, functionally related genes are usually limited to specific compartments for efficient cascade gene expression For example, in eukaryotic cells, chromosomes are divided into several chromosomal domains according to the correlation of gene function; a series of genes encoding respiratory chain complex are arranged on mitochondrial DNA and wrapped in mitochondria Therefore, the study of the effect of DNA compartmentalization on cascade gene expression can provide important enlightenment for understanding the relationship between cell structure and function Recently, Yang Dayong, Professor of the school of chemical engineering of Tianjin University, constructed the gene circuit compartmentalization structure on the nano interface by anchoring two functionally related genes on the gold nanoparticles, and studied the effect of the gene circuit compartmentalization on the cascade gene expression by using the cell-free gene expression system In this study, the model gene line is composed of T7 RNA polymerase gene and reporter protein enhanced green fluorescent protein gene A dendrimer with two mercapto modified scaffolds was assembled at one end of each gene by polymerase chain reaction (PCR) The two genes were anchored on the surface of gold nanoparticles by gold sulfur bond, and the gene circuit compartment structure was constructed on the nano interface The route compartmentalization structure of the gene brings the physical distance between the regulatory protein gene and the reporter protein gene closer to the nanometer scale, which makes the synthesized regulatory protein RNA polymerase mainly distributed in the region of the compartmentalization and quickly shuttle to the adjacent reporter protein gene, thus promoting the cascade gene expression The experimental results showed that the compartmentalized structure of gene circuit increased the yield and initial expression rate of reporter protein, and shortened the feedback time of cascade gene expression In addition, this kind of gene line compartmentalization effect is dependent on gene concentration: compared with the high gene concentration, the gene line compartmentalization structure can play a more spatial compartmentalization effect in a relatively low gene concentration, and significantly improve the efficiency of cascade gene expression This work not only provides a simplified model for constructing gene circuit compartmentalization on nano interface, but also provides important enlightenment for understanding the relationship between cell construction and function The research results were published in Journal of the American Chemical Society (DOI: 10.1021 / JACS 9b11407), and relevant achievements have applied for Chinese invention patents Guo Xiaocui and Li Feng, Ph.D students, School of chemical engineering, Tianjin University, are co authors The research is supported by NSFC.