Amazing stuff!
"... a new tool that can light up any region of the genome in any living cell and enable scientists to watch how different regions interact with one another. ..."
From the highlights and abstract:
"Highlights
• Oligo-LiveFISH tracks non-repetitive loci in broad cell types and primary cells
• High spatial (20 nm) and temporal (50 ms) resolution chromatin imaging in live cells
• Dynamic tracking and modeling reveal 1D cis- and 3D trans-communication of genomic DNA
• Active transcription induces enhancer and promoter confinement and slower dynamics
Summary
Three-dimensional (3D) genome dynamics are crucial for cellular functions and disease. However, real-time, live-cell DNA visualization remains challenging, as existing methods are often confined to repetitive regions, suffer from low resolution, or require complex genome engineering.
Here, we present Oligo-LiveFISH, a high-resolution, reagent-based platform for dynamically tracking non-repetitive genomic loci in diverse cell types, including primary cells. Oligo-LiveFISH utilizes fluorescent guide RNA (gRNA) oligo pools generated by computational design, in vitro transcription, and chemical labeling, delivered as ribonucleoproteins.
Utilizing machine learning, we characterized the impact of gRNA design and chromatin features on imaging efficiency.
Multi-color Oligo-LiveFISH achieved 20-nm spatial resolution and 50-ms temporal resolution in 3D, capturing real-time enhancer and promoter dynamics. Our measurements and dynamic modeling revealed two distinct modes of chromatin communication, and active transcription slows enhancer-promoter dynamics at endogenous genes like FOS.
Oligo-LiveFISH offers a versatile platform for studying 3D genome dynamics and their links to cellular processes and disease."
High-resolution dynamic imaging of chromatin DNA communication using Oligo-LiveFISH (no public access)
Graphical abstract
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