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从活细胞中的单端粒表达的癌症细胞克隆的生成, 以可视化含有端粒重复的 rna terra

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Generation of Cancer Cell Clones to Visualize Telomeric Repeat-containing RNA TERRA Expressed from a Single Telomere in Living Cells

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从活细胞中的单端粒表达的癌症细胞克隆的生成, 以可视化含有端粒重复的 rna terra

DOI:

10.3791/58790-v

•

09:13 min

•

January 17, 2019

•

Laura Avogaro, Claudio Oss Pegorar, Nicole Bettin², Emilio Cusanelli

¹Laboratory of Cell Biology and Molecular Genetics, Department of Cellular, Computational and Integrative Biology – CIBIO,University of Trento, ²Department of Life Sciences,University of Trieste

Chapters

00:04Title
00:37Selection of Neomycin Resistant Clones
03:41Screening of Neomycin Resistant Clones
05:34Elimination of the Neomycin Resistance Gene from the MS2 Cassette Integrated at Subtelomere 15q
06:06Visualization of TERRA-MS2 Transcripts in Living Cells
06:49Results: Cancer Cell Clones with MS2 Sequence Tag Created and Imaged
08:17Conclusion

Summary

자동 번역

在这里, 我们提出了一个协议, 以生成包含 ms2 序列标记在单个亚端粒的癌细胞克隆。这种方法, 依靠 ms2-gfp 系统, 实现了端粒重复 rna (terra) 的内源性转录记录的可视化表达从一个端粒表达在活细胞中。

Tags

Keywords: Cancer Cell Clones Telomeric Repeat-containing RNA (TERRA)Yeast Telomeric Noncoding RNA Telomere Cell Visualization Single Telomere Cell Culture CRISPR-Cas9 Cell Cloning Cell Freezing Fluorescence Microscopy

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