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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
Este relatório descreve um método de bioengenharia para conceber e construir novos factores de splicing artificial (ASFS) que modulam especificamente a splicing de genes alvo em células de mamífero. Este método pode ser expandido para manipular vários fatores artificiais para manipular outros aspectos do metabolismo do ARN.
