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基于细胞的 sineup 非编码 rna 检测, 可专门增强 mrna 的翻译能力
JoVE Journal
Genetics
This content is Free Access.
JoVE Journal Genetics
Cell Based Assays of SINEUP Non-coding RNAs That Can Specifically Enhance mRNA Translation
DOI:

10:21 min

February 01, 2019

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Chapters

  • 00:04Title
  • 00:48Design of Binding Domains (BDs) of SINEUP Constructs for Target mRNAs
  • 01:48Cell Culture and SINEUPs Transfection
  • 03:46Protein Extraction
  • 05:32Protein Separation by SDS-PAGE and Detection of Target Proteins by Western-blot Analysis
  • 07:21Imaging Analysis of SINEUPs by Semi-automated Detection Method
  • 08:26Results: Analysis of Translation Upregulation by SINEUP-GFP
  • 09:11Conclusion

Summary

Automatic Translation

sineup 是一种合成反义非编码 rna, 它包含一个结合域 (bd) 和一个效应域 (ed), 并向上调节目标 mrna 的翻译。在这里, 我们描述了在培养细胞系中检测 sineup 的方法, 分析了他们的翻译促进活性的西方印迹和半自动高通量成像系统。

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