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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
JoVE Journal
Genetics
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JoVE Journal Genetics
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

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09:13 min

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January 17, 2019

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, , ,
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

Automatic Translation

Automatic Translation

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

Cancer Cell ClonesTelomeric Repeat-containing RNA (TERRA)Yeast Telomeric Noncoding RNATelomereCell VisualizationSingle TelomereCell CultureCRISPR-Cas9Cell CloningCell FreezingFluorescence Microscopy

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