CAS OpenIR  > 中科院上海应用物理研究所2011-2020年
Genetically encoded X-ray cellular imaging for nanoscale protein localization
Kong, HT; Zhang, JC; Li, J; Wang, J; Shin, HJ; Tai, RZ; Yan, QL; Xia, K; Hu, J; Wang, LH; Zhu, Y; Fan, CH
AbstractSpatial resolution defines the physical limit of microscopes for probing biomolecular localization and interactions in cells. Whereas synchrotron-based X-ray microscopy (XRM) represents a unique approach for imaging a whole cell with nanoscale resolution due to its intrinsic nanoscale resolution and great penetration ability, existing approaches to label biomolecules rely on the use of exogenous tags that are multi-step and error-prone. Here, we repurpose engineered peroxidases as genetically encoded X-ray-sensitive tags (GXET) for site-specific labeling of protein-of-interest in mammalian cells. We find that 3,3'-diaminobenzidine (DAB) polymers that are in-situ catalytically formed by fusion-expressed peroxidases are visible under XRM. Using this new tag, we imaged the protein location associated with the alteration of a DNA-methylation pathway with an ultra-high resolution of 30 nanometers. Importantly, the excellent energy resolution of XRM enables multicolor imaging using different peroxidase tags. The development of GXET enlightens the way to nanoscopic imaging for biological studies.
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Affiliation1.Chinese Acad Sci, Shanghai Adv Res Inst, Bioimaging Ctr, Shanghai 201210, Peoples R China
2.Chinese Acad Sci, Shanghai Inst Appl Phys, Div Phys Biol, CAS Key Lab Interfacial Phys & Technol, Shanghai 201800, Peoples R China
3.Univ Saskatchewan, Canadian Light Source Inc, Saskatoon, SK S7N 2V3, Canada
4.POSTECH, Pohang Accelerator Lab, Pohang 37673, South Korea
5.Shanghai Jiao Tong Univ, Sch Chem & Chem Engn, Frontiers Sci Ctr Transformat Mol, Shanghai 200240, Peoples R China
6.Shanghai Jiao Tong Univ, Shanghai Key Lab Nucle Acids Chem & Nanomed, Inst Mol Med, Renji Hosp,Sch Med, Shanghai 200240, Peoples R China
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GB/T 7714
Kong, HT,Zhang, JC,Li, J,et al. Genetically encoded X-ray cellular imaging for nanoscale protein localization[J]. NATIONAL SCIENCE REVIEW,2020,7(7):1218-1227.
APA Kong, HT.,Zhang, JC.,Li, J.,Wang, J.,Shin, HJ.,...&Fan, CH.(2020).Genetically encoded X-ray cellular imaging for nanoscale protein localization.NATIONAL SCIENCE REVIEW,7(7),1218-1227.
MLA Kong, HT,et al."Genetically encoded X-ray cellular imaging for nanoscale protein localization".NATIONAL SCIENCE REVIEW 7.7(2020):1218-1227.
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