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Cell Based Assays of SINEUP Non-coding RNAs That Can Specifically Enhance mRNA Translation
JoVE Journal
Genética
This content is Free Access.
JoVE Journal Genética
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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Capítulos

  • 00:04Título
  • 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

Tadução automática

SINEUPs are synthetic antisense non-coding RNAs, which contain a binding domain (BD) and an effector domain (ED) and up-regulate translation of target mRNA. Here, we describe detection methods for SINEUPs in cultured cell lines, analysis of their translation-promoting activity by Western-blot and a semi-automated high throughput imaging system.

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