Journal
/
Genética
/
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
Genética
É necessária uma assinatura da JoVE para visualizar este conteúdo.  Faça login ou comece sua avaliação gratuita.
JoVE Journal Genética
Generation of Cancer Cell Clones to Visualize Telomeric Repeat-containing RNA TERRA Expressed from a Single Telomere in Living Cells
Please note that all translations are automatically generated. Click here for the English version.

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

•

, , ,
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

Capítulos

  • 00:04Título
  • 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

Tadução automática

Tadução automática

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

Article

Embed

ADICIONAR À PLAYLIST

Usage Statistics

Vídeos Relacionados

工程人为因素来特异性地操纵人细胞中的选择性剪接

饲养并在隐藏甲壳虫双链RNA介导的基因敲除，<em>白腹皮蠹</em

单细胞测序拷贝数改变的检测

单细胞基因表达谱使用流式细胞仪和qPCR内部标准

使用生物素化补骨脂素全球定位RNA-RNA相互作用

用荧光PCR毛细管电泳技术在基因型CRISPR / Cas9介导的敲除突变体的高通量形式

选择依赖和独立生成CRISPR / Cas9介导的哺乳动物细胞基因敲除

人端粒酶逆转录蛋白 rna 依赖性 rna 聚合酶活性的半定量检测

基于细菌选择和生长的盘状细胞基因工程

从前列腺癌的遗传工程小鼠模型生成肿瘤有机体

Read Article