远红色化学遗传激酶生物传感器实现信号网络的多路复用和超分辨率成像

IF 33.1 1区 生物学 Q1 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Michelle S. Frei, Samantha A. Sanchez, Xinchang He, Longwei Liu, Falk Schneider, Zichen Wang, Hiroyuki Hakozaki, Yajuan Li, Anne C. Lyons, Theresa V. Rohm, Jerrold M. Olefsky, Lingyan Shi, Johannes Schöneberg, Scott E. Fraser, Sohum Mehta, Yingxiao Wang, Jin Zhang
{"title":"远红色化学遗传激酶生物传感器实现信号网络的多路复用和超分辨率成像","authors":"Michelle S. Frei, Samantha A. Sanchez, Xinchang He, Longwei Liu, Falk Schneider, Zichen Wang, Hiroyuki Hakozaki, Yajuan Li, Anne C. Lyons, Theresa V. Rohm, Jerrold M. Olefsky, Lingyan Shi, Johannes Schöneberg, Scott E. Fraser, Sohum Mehta, Yingxiao Wang, Jin Zhang","doi":"10.1038/s41587-025-02642-8","DOIUrl":null,"url":null,"abstract":"<p>Fluorescent biosensors have advanced biomedical research by enabling direct live-cell measurements of signaling activities. However, current technology offers limited resolution and dimensionality, impeding our ability to resolve and interrogate spatiotemporally regulated networks of signaling activities. Here we introduce highly sensitive chemigenetic kinase activity biosensors that combine the genetically encodable self-labeling tag, HaloTag7, with far-red-emitting synthetic fluorophores. This technology enables both four-dimensional activity imaging and functional super-resolution imaging using stimulated emission depletion and other high-resolution microscopy techniques, permitting signaling activity to be detected across scales with the necessary resolution. Stimulated emission depletion imaging enabled the investigation of protein kinase A activity at individual clathrin-coated pits. We further demonstrate imaging of up to five analytes in single living cells, an increase in the dimensionality of biosensor multiplexing. Multiplexed imaging of cellular responses to the activation of different G-protein-coupled receptors (GPCRs) allowed quantitative measurements of spatiotemporal network states downstream of individual GPCR–ligand pairs.</p>","PeriodicalId":19084,"journal":{"name":"Nature biotechnology","volume":"14 1","pages":""},"PeriodicalIF":33.1000,"publicationDate":"2025-04-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Far-red chemigenetic kinase biosensors enable multiplexed and super-resolved imaging of signaling networks\",\"authors\":\"Michelle S. Frei, Samantha A. Sanchez, Xinchang He, Longwei Liu, Falk Schneider, Zichen Wang, Hiroyuki Hakozaki, Yajuan Li, Anne C. Lyons, Theresa V. Rohm, Jerrold M. Olefsky, Lingyan Shi, Johannes Schöneberg, Scott E. Fraser, Sohum Mehta, Yingxiao Wang, Jin Zhang\",\"doi\":\"10.1038/s41587-025-02642-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Fluorescent biosensors have advanced biomedical research by enabling direct live-cell measurements of signaling activities. However, current technology offers limited resolution and dimensionality, impeding our ability to resolve and interrogate spatiotemporally regulated networks of signaling activities. Here we introduce highly sensitive chemigenetic kinase activity biosensors that combine the genetically encodable self-labeling tag, HaloTag7, with far-red-emitting synthetic fluorophores. This technology enables both four-dimensional activity imaging and functional super-resolution imaging using stimulated emission depletion and other high-resolution microscopy techniques, permitting signaling activity to be detected across scales with the necessary resolution. Stimulated emission depletion imaging enabled the investigation of protein kinase A activity at individual clathrin-coated pits. We further demonstrate imaging of up to five analytes in single living cells, an increase in the dimensionality of biosensor multiplexing. Multiplexed imaging of cellular responses to the activation of different G-protein-coupled receptors (GPCRs) allowed quantitative measurements of spatiotemporal network states downstream of individual GPCR–ligand pairs.</p>\",\"PeriodicalId\":19084,\"journal\":{\"name\":\"Nature biotechnology\",\"volume\":\"14 1\",\"pages\":\"\"},\"PeriodicalIF\":33.1000,\"publicationDate\":\"2025-04-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Nature biotechnology\",\"FirstCategoryId\":\"5\",\"ListUrlMain\":\"https://doi.org/10.1038/s41587-025-02642-8\",\"RegionNum\":1,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Nature biotechnology","FirstCategoryId":"5","ListUrlMain":"https://doi.org/10.1038/s41587-025-02642-8","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

荧光生物传感器通过直接测量信号活动的活细胞,促进了生物医学研究。然而,目前的技术提供了有限的分辨率和维度,阻碍了我们解决和询问时空调节的信号活动网络的能力。在这里,我们介绍了高度敏感的化学遗传激酶活性生物传感器,它结合了遗传可编码的自标记标签HaloTag7和远红色合成荧光团。该技术利用受激发射损耗和其他高分辨率显微镜技术,实现了四维活动成像和功能超分辨率成像,从而可以在必要的分辨率下跨尺度检测信号活动。受激发射耗竭成像使研究蛋白激酶A活性在单个网格蛋白包被凹坑。我们进一步展示了在单个活细胞中多达五种分析物的成像,增加了生物传感器多路复用的维度。细胞对不同g蛋白偶联受体(gpcr)激活反应的多重成像允许定量测量单个gpcr配体对下游的时空网络状态。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Far-red chemigenetic kinase biosensors enable multiplexed and super-resolved imaging of signaling networks

Far-red chemigenetic kinase biosensors enable multiplexed and super-resolved imaging of signaling networks

Fluorescent biosensors have advanced biomedical research by enabling direct live-cell measurements of signaling activities. However, current technology offers limited resolution and dimensionality, impeding our ability to resolve and interrogate spatiotemporally regulated networks of signaling activities. Here we introduce highly sensitive chemigenetic kinase activity biosensors that combine the genetically encodable self-labeling tag, HaloTag7, with far-red-emitting synthetic fluorophores. This technology enables both four-dimensional activity imaging and functional super-resolution imaging using stimulated emission depletion and other high-resolution microscopy techniques, permitting signaling activity to be detected across scales with the necessary resolution. Stimulated emission depletion imaging enabled the investigation of protein kinase A activity at individual clathrin-coated pits. We further demonstrate imaging of up to five analytes in single living cells, an increase in the dimensionality of biosensor multiplexing. Multiplexed imaging of cellular responses to the activation of different G-protein-coupled receptors (GPCRs) allowed quantitative measurements of spatiotemporal network states downstream of individual GPCR–ligand pairs.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Nature biotechnology
Nature biotechnology 工程技术-生物工程与应用微生物
CiteScore
63.00
自引率
1.70%
发文量
382
审稿时长
3 months
期刊介绍: Nature Biotechnology is a monthly journal that focuses on the science and business of biotechnology. It covers a wide range of topics including technology/methodology advancements in the biological, biomedical, agricultural, and environmental sciences. The journal also explores the commercial, political, ethical, legal, and societal aspects of this research. The journal serves researchers by providing peer-reviewed research papers in the field of biotechnology. It also serves the business community by delivering news about research developments. This approach ensures that both the scientific and business communities are well-informed and able to stay up-to-date on the latest advancements and opportunities in the field. Some key areas of interest in which the journal actively seeks research papers include molecular engineering of nucleic acids and proteins, molecular therapy, large-scale biology, computational biology, regenerative medicine, imaging technology, analytical biotechnology, applied immunology, food and agricultural biotechnology, and environmental biotechnology. In summary, Nature Biotechnology is a comprehensive journal that covers both the scientific and business aspects of biotechnology. It strives to provide researchers with valuable research papers and news while also delivering important scientific advancements to the business community.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信