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Full-Text Articles in Genetics and Genomics
Genome Engineering In Mammalian Cells By Flp And Cre Dna Recombinase Variants, Riddhi Shah
Genome Engineering In Mammalian Cells By Flp And Cre Dna Recombinase Variants, Riddhi Shah
Doctoral Dissertations
Genome engineering relies on DNA modifying enzymes that are able to locate a DNA sequence of interest and initiate a desired genome rearrangement. Currently, the field predominantly utilizes site-specific DNA nucleases that depend on the host DNA repair machinery to complete a genome modification task. We show here that genome engineering approaches that employ self-sufficient, versatile site-specific DNA recombinase Flp and Cre can be developed into promising alternatives. We demonstrate that a Flp variant evolved to recombine an FRT-like sequence FL-IL10A, which is located upstream of the human interleukin-10 gene, can target this sequence in the model setting and native …
Size Specific Transfection To Mammalian Cells By Micropillar Array Electroporation, Yingbo Zu
Size Specific Transfection To Mammalian Cells By Micropillar Array Electroporation, Yingbo Zu
Doctoral Dissertations
Electroporation serves as a promising non-viral gene delivery approach, while its current configurations carry drawbacks associated with high-voltage electrical pulses and heterogeneous treatment on individual cells. Here, we developed a new micropillar array electroporation (MAE) platform to advance the delivery of plasmid DNA and RNA to mammalian cells. By introducing well-patterned micropillar array on the electrode surface, the number of pillars each cell faces varies with its cell membrane surface area, despite their large population and random locations. In this way, cell size specific electroporation is conveniently done and contributed to a 2.5~3 fold increase on plasmid DNA transfection and …