BRIGHT Enables High-SNR Live-Cell Imaging of Non-Repetitive Sequences via Bivalent Fluorescent Nanobody-Mediated Cascade-Dependent Illumination.

IF 14.1 1区材料科学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Advanced Science Pub Date : 2025-10-17 DOI:10.1002/advs.202513014

Lei Feng, Tao Huang, Yanxi Han, Meng Tian, Duo Wang, Zilong Mei, Yu Ma, Yingshuo Ma, Jinming Li, Rui Zhang

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引用次数: 0

Abstract

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_ALFA). 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.

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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。

基因组位点和基因组无关DNA表现出与生理功能相关的异质性动态。非重复序列的活细胞成像是必要的，但受低信噪比（SNR）的限制，限制了准确识别和动态跟踪。本文报道了一种用于非重复序列实时成像的BRIGHT（二价近红外纳米体介导的基因组位点级联照明，用于高信噪比跟踪）系统的开发。BRIGHT利用dCas9-n×ALFA靶向基因组位点，并通过双价结合和抗原依赖照明触发级联依赖性照明，实现高信噪比靶向ALFA标签的二价近红外荧光纳米体（Bi-NIR-FbALFA）。BRIGHT在端粒标记上的信噪比分别比SunTag和PP7-PCP高≈6.26倍和≈6.76倍，实现非重复DNA成像，信噪比高达284.84。此外，与以前的工具相比，BRIGHT可以跟踪染色质动力学，将动力学和核定位与转录活性联系起来，并检测染色体外环状DNA （eccDNA）的分布和动态异质性。总体而言，BRIGHT显著提高了非重复序列成像的信噪比，为研究基因组动力学和基因调控提供了一种创新工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Advanced Science CHEMISTRY, MULTIDISCIPLINARYNANOSCIENCE &-NANOSCIENCE & NANOTECHNOLOGY

CiteScore

18.90

自引率

2.60%

发文量

1602

审稿时长

1.9 months

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.