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SJTU Researchers Published Research Article in Advanced Materials

May 18, 2021      Author:

Recently, SJTU Professor Ding Xianting and Assistant Researcher Xie Haiyang published an article in Advanced Materials, reporting a single‐cell transfection analysis chip (scTAC) based on the in situ microchip immunoblotting method for rapid and accurate analysis of exogenous gene expression in thousands of individual host cells. scTAC not only provides sensitive, rapid and accurate analysis of information on exogenous gene expression activity in individual cells, but also can be used for the analysis of transfection efficiency of rare cells and evaluation of transfection methods.

This is an innovative application research on single cell immunoblot analysis technology in the field of cell molecular biology, which effectively solves the difficulties in transfection efficiency analysis and transfection technology research in rare cells. The technology realizes the localization, screening and detection of transfected cells, and the whole analysis process can be completed within 4 hours. At the same time, the chip is small and portable, simple to store, and the back-end detection equipment can be replaced as needed, demonstrating great application potentials in cell molecular biology research and clinical medical diagnosis.

Doctoral student Li Shanhe is the first author of the paper, and Professor Ding Xianting and Assistant Researcher Xie Haiyang are the corresponding authors. The related research was supported by the National Natural Science Foundation of China, the National Key Research and Development Program, and the Shanghai Science and Technology Innovation Initiative.

Article Link: Li, S. et al. Single-Cell Immunoblotting based on a Photoclick Hydrogel Enables High-Throughput Screening and Accurate Profiling of Exogenous Gene Expression. Advanced Materials n/a, 2101108, doi:https://doi.org/10.1002/adma.202101108 (2021).

 

Author: Yufang Xin

Source: Institute of Personalized Medicine, SJTU

Translated by Zhang Yue

Proofread by Xiao Yangning, Fu Yuhe

 

ABSTRACT:

Fast and accurate profiling of exogenous gene expression in host cells is crucial for studying gene function in cellular and molecular biology, but still faces the challenge of incomplete co‐expression of reporter genes and target genes. Here, a single‐cell transfection analysis chip (scTAC) is presented, which is based on the in situ microchip immunoblotting method, for rapid and accurate analysis of exogenous gene expression in thousands of individual host cells. scTAC not only can assign information of exogenous gene activity to specific transfected cells, but enables the acquisition of continuous protein expression even in low co‐expression scenarios. It is demonstrated that scTAC can reveal the relationship of expression level between reporter genes and target genes, which is helpful for evaluating transient transfection strategy efficiency. The advantages of this method for the study of fusion protein expression and downstream protein expression in signaling pathway in rare cells are shown. Empirically, an EGFP‐TSPAN8 fusion plasmid is transfected into MCF‐7 breast cancer cells and the expressions of two cancer stemness biomarkers (ALDHA1 and SOX2) are analyzed. The scTAC method clearly reveals an interesting phenomenon that transfected adherent MCF‐7 cells exhibit some stem cell characteristics, but they do not have stem cell appearance.