Lei Feng, Tao Huang, Yanxi Han, Meng Tian, Duo Wang, Zilong Mei, Yu Ma, Yingshuo Ma, Jinming Li, Rui Zhang
{"title":"BRIGHT通过二价荧光纳米体介导的级联依赖照明实现非重复序列的高信噪比活细胞成像。","authors":"Lei Feng, Tao Huang, Yanxi Han, Meng Tian, Duo Wang, Zilong Mei, Yu Ma, Yingshuo Ma, Jinming Li, Rui Zhang","doi":"10.1002/advs.202513014","DOIUrl":null,"url":null,"abstract":"<p><p>Genomic loci and genome-independent DNA exhibit heterogeneous dynamics related to physiological function. Live-cell imaging of non-repetitive sequences is essential but limited by low signal-to-noise ratio (SNR), restricting accurate identification and dynamic tracking. Here, the development of a BRIGHT (bivalent near-infrared nanobody-mediated cascade illumination of genomic loci for high-SNR tracking) system for non-repetitive sequence live imaging is reported. BRIGHT employs dCas9-n×ALFA to target genomic loci and realizes high SNR by triggering cascade-dependent illumination via bivalent binding and antigen-dependent illumination of a bivalent near-infrared fluorescent nanobody targeting ALFA tags (Bi-NIR-Fb<sub>ALFA</sub>). BRIGHT achieves ≈6.26-fold and ≈6.76-fold higher SNR than SunTag and PP7-PCP in telomere labeling, and enables non-repetitive DNA imaging with SNR up to 284.84. Furthermore, BRIGHT tracks chromatin dynamics comparably to previous tools, links dynamics and nuclear positioning to transcriptional activity, and detects the distribution and dynamic heterogeneity of extrachromosomal circular DNA (eccDNA). Overall, BRIGHT dramatically improves the SNR of non-repetitive sequence imaging and offers an innovative tool for studying genomic dynamics and gene regulation.</p>","PeriodicalId":117,"journal":{"name":"Advanced Science","volume":" ","pages":"e13014"},"PeriodicalIF":14.1000,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"BRIGHT Enables High-SNR Live-Cell Imaging of Non-Repetitive Sequences via Bivalent Fluorescent Nanobody-Mediated Cascade-Dependent Illumination.\",\"authors\":\"Lei Feng, Tao Huang, Yanxi Han, Meng Tian, Duo Wang, Zilong Mei, Yu Ma, Yingshuo Ma, Jinming Li, Rui Zhang\",\"doi\":\"10.1002/advs.202513014\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Genomic loci and genome-independent DNA exhibit heterogeneous dynamics related to physiological function. Live-cell imaging of non-repetitive sequences is essential but limited by low signal-to-noise ratio (SNR), restricting accurate identification and dynamic tracking. Here, the development of a BRIGHT (bivalent near-infrared nanobody-mediated cascade illumination of genomic loci for high-SNR tracking) system for non-repetitive sequence live imaging is reported. BRIGHT employs dCas9-n×ALFA to target genomic loci and realizes high SNR by triggering cascade-dependent illumination via bivalent binding and antigen-dependent illumination of a bivalent near-infrared fluorescent nanobody targeting ALFA tags (Bi-NIR-Fb<sub>ALFA</sub>). BRIGHT achieves ≈6.26-fold and ≈6.76-fold higher SNR than SunTag and PP7-PCP in telomere labeling, and enables non-repetitive DNA imaging with SNR up to 284.84. Furthermore, BRIGHT tracks chromatin dynamics comparably to previous tools, links dynamics and nuclear positioning to transcriptional activity, and detects the distribution and dynamic heterogeneity of extrachromosomal circular DNA (eccDNA). Overall, BRIGHT dramatically improves the SNR of non-repetitive sequence imaging and offers an innovative tool for studying genomic dynamics and gene regulation.</p>\",\"PeriodicalId\":117,\"journal\":{\"name\":\"Advanced Science\",\"volume\":\" \",\"pages\":\"e13014\"},\"PeriodicalIF\":14.1000,\"publicationDate\":\"2025-10-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Advanced Science\",\"FirstCategoryId\":\"88\",\"ListUrlMain\":\"https://doi.org/10.1002/advs.202513014\",\"RegionNum\":1,\"RegionCategory\":\"材料科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"CHEMISTRY, MULTIDISCIPLINARY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Advanced Science","FirstCategoryId":"88","ListUrlMain":"https://doi.org/10.1002/advs.202513014","RegionNum":1,"RegionCategory":"材料科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
BRIGHT Enables High-SNR Live-Cell Imaging of Non-Repetitive Sequences via Bivalent Fluorescent Nanobody-Mediated Cascade-Dependent Illumination.
Genomic loci and genome-independent DNA exhibit heterogeneous dynamics related to physiological function. Live-cell imaging of non-repetitive sequences is essential but limited by low signal-to-noise ratio (SNR), restricting accurate identification and dynamic tracking. Here, the development of a BRIGHT (bivalent near-infrared nanobody-mediated cascade illumination of genomic loci for high-SNR tracking) system for non-repetitive sequence live imaging is reported. BRIGHT employs dCas9-n×ALFA to target genomic loci and realizes high SNR by triggering cascade-dependent illumination via bivalent binding and antigen-dependent illumination of a bivalent near-infrared fluorescent nanobody targeting ALFA tags (Bi-NIR-FbALFA). BRIGHT achieves ≈6.26-fold and ≈6.76-fold higher SNR than SunTag and PP7-PCP in telomere labeling, and enables non-repetitive DNA imaging with SNR up to 284.84. Furthermore, BRIGHT tracks chromatin dynamics comparably to previous tools, links dynamics and nuclear positioning to transcriptional activity, and detects the distribution and dynamic heterogeneity of extrachromosomal circular DNA (eccDNA). Overall, BRIGHT dramatically improves the SNR of non-repetitive sequence imaging and offers an innovative tool for studying genomic dynamics and gene regulation.
期刊介绍：
Advanced Science is a prestigious open access journal that focuses on interdisciplinary research in materials science, physics, chemistry, medical and life sciences, and engineering. The journal aims to promote cutting-edge research by employing a rigorous and impartial review process. It is committed to presenting research articles with the highest quality production standards, ensuring maximum accessibility of top scientific findings. With its vibrant and innovative publication platform, Advanced Science seeks to revolutionize the dissemination and organization of scientific knowledge.
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