Computer-Aided Design of DNA Self-Limited Assembly for Relative Quantification of Membrane Proteins
Computer-Aided Design of DNA Self-Limited Assembly for Relative Quantification of Membrane Proteins
复制标题
用于膜蛋白相对定量的 DNA 自限制组装的计算机辅助设计
DOI：
10.1021/acs.analchem.2c01909
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发表时间：
2022
影响因子：
7.4
通讯作者：
Weihong Tan
中科院分区：
文献类型：
--
作者：
Dongsheng Mao;Cuicui Lu;Runchi Zhang;Liucun Zhu;Yuchen Song;Chang Feng;Qianqian Zhang;Tianshu Chen;Yu Yang;Guifang Chen;Xiaoli Zhu;Weihong Tan
Immunofluorescence imaging of cells plays a vital role in biomedical research and clinical diagnosis. However, when it is applied to relative quantification of proteins, it suffers from insufficient fluorescence intensity or partial overexposure, resulting in inaccurate relative quantification. Herein, we report a computer-aided design of DNA self-limited assembly (CAD-SLA) technology and apply it for relative quantification of membrane proteins, a concept proposed for the first time. CAD-SLA can achieve exponential cascade signal amplification in one pot and terminate at any desired level. By conjugating CAD-SLA with immunofluorescence, in situ imaging of cell membrane proteins is achieved with a controllable amplification level. Besides, comprehensive fluorescence intensity information from fluorescent images can be obtained, accurately showing relative quantitative information. Slight protein expression differences previously indistinguishable by immunofluorescence or Western blotting can now be discriminated, making fluorescence imaging-based relative quantification a promising tool for membrane protein analysis. From the perspectives of both DNA self-assembly technology and immunofluorescence technology, this work has solved difficult problems and provided important reference for future development.
