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Direct synthesis of EM-visible gold nanoparticles on genetically encoded tags for single-molecule visualization in cells

By Zhaodi Jiang, Xiumei Jin, Yuhua Li, Sitong Liu, Xiao-Man Liu, Ying-Ying Wang, Pei Zhao, Xinbin Cai, Ying Liu, Yaqi Tang, Xiaobin Sun, Yan Liu, Yanyong Hu, Ming Li, Gaihong Cai, Xiangbing Qi, She Chen, Li-Lin Du, Wanzhong He

Posted 17 Jan 2019
bioRxiv DOI: 10.1101/520999

Single-molecule visualization in cells with genetically encoded tags for electron microscopy (EM) has been a long-awaited but unimplemented tool for cell biologists. Here, we report an approach for directly synthesizing EM-visible gold nanoparticles (AuNPs) on cysteine-rich tags for single-molecule visualization in cells. We first uncovered an auto-nucleation suppression mechanism that allows specific synthesis of AuNPs on isolated cysteine-rich tags. We next exploited this mechanism to develop an approach for single-molecule detection of proteins in prokaryotic cells and achieved an unprecedented labeling efficiency. We then expanded it to more complicated eukaryotic cells and successfully detected the proteins targeted to various organelles, including the membranes of endoplasmic reticulum (ER) and nuclear envelope, ER lumen, nuclear pores, spindle pole bodies, and mitochondrial matrix. Thus, our implementation of genetically encoded tags for EM should allow cell biologists to address an enormous range of biological questions at single-molecule level in diverse cellular ultrastructural contexts without using antibodies.

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