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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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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

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

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

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Here, we present a protocol to generate cancer cell clones containing a MS2 sequence tag at a single subtelomere. This approach, relying on the MS2-GFP system, enables visualization of the endogenous transcripts of telomeric repeat-containing RNA (TERRA) expressed from a single telomere in living cells.

Tags

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

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