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工程人为因素来特异性地操纵人细胞中的选择性剪接
JoVE Journal
Genetics
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JoVE Journal Genetics
Engineering Artificial Factors to Specifically Manipulate Alternative Splicing in Human Cells
DOI:

10:06 min

April 26, 2017

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Chapters

  • 00:05Title
  • 00:51Construction of the PUF RNA-binding Domain Scaffold with Customized RNA Binding Specificity by Overlapping PCR
  • 02:05Transfection of Engineered Splicing Factors (ESFs) for Modulation of Exon Inclusion and Alternative Splice-site Use
  • 03:43Cell Harvest and Identification of Spliced Isoforms
  • 06:24Modulation of Endogenous Bcl-x Splicing and Apoptosis by ESFs
  • 08:23Results: Subcellular Localization of ESFs in Transfected HeLa Cells
  • 09:30Conclusion

Summary

Automatic Translation

该报告介绍了一种生物工程方法来设计和构建特异性调节在哺乳动物细胞中靶基因的剪接新颖人工剪接因子(ASFS)。该方法可以进一步扩展来设计各种人为因素来操纵RNA代谢的其它方面。

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