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通过Oligo-LiveFISH可视化活细胞中的染色体通信

Mattia Conte¹, Mario Nicodemi¹

¹Dipartimento di Fisica, Università di Napoli Federico II and INFN Napoli, Complesso Universitario di Monte Sant'Angelo, 80126 Naples, Italy.

Cell reports methods

|June 10, 2025

概括

研究人员开发了Oligo-LiveFISH,这是一种新的活细胞成像技术. 这种方法以高分辨率跟踪基因组组织动态,而不需要进行遗传修饰.

科学领域:

分子生物学分子生物学
遗传学是一个遗传学.
细胞生物学细胞生物学

背景情况:

了解基因组调节需要实时观察染色质动态.
目前用于追踪活细胞中的基因组位置的方法通常需要基因组工程,从而限制了它们的应用.

研究的目的:

推出Oligo-LiveFISH,这是一个新的平台,用于高分辨率的染色体动态的活细胞成像.
为了能够在不需要基因组工程的情况下跟踪非重复的基因组位置.

主要方法:

组装光导向RNA池,使用催化失活的Cas9 (dCas9).
利用组装的复合体对活细胞中的特定基因组位点进行高分辨率成像.
实现20纳米的空间分辨率和50毫秒的时间分辨率.

主要成果:

演示了Oligo-LiveFISH在实时跟踪非重复性基因组位点的能力.
在各种细胞系和初级细胞中成功应用了该平台.
提供了对基因组动态组织的见解.

结论:

奥利戈-LiveFISH为研究基因组调节和动态提供了一个强大的,非侵入性的工具.
该平台的高分辨率和多功能性推动了基因组研究活细胞成像领域的发展.
这种方法有助于更深入地了解不同细胞环境中的动态基因组组织.

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FISH - Fluorescent In-situ Hybridization

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...