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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
This report describes a bioengineering method to design and construct novel Artificial Splicing Factors (ASFs) that specifically modulate the splicing of target genes in mammalian cells. This method can be further expanded to engineer various artificial factors to manipulate other aspects of RNA metabolism.
