/

/

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
Genetica

È necessario avere un abbonamento a JoVE per visualizzare questo. Accedi o inizia la tua prova gratuita.

JoVE Journal Genetica

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

•

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

Capitoli

00:04Titolo
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

Traduzione automatica

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

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

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