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The miR-155* targeting TALEN pair causes both bi-allelic and mono-allelic mutations in human cells.

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posted on 07.05.2013 by Ruozhen Hu, Jared Wallace, Timothy J. Dahlem, David Jonah Grunwald, Ryan M. O'Connell

(A) Schematic of the experimental design. 293T cells were transfected with the TALEN pair targeting the miR-155* region along with a GFP plasmid. 48 hours later, GFP+ cells were sorted by FACS and subjected to single cell cloning. After individual cell clones were expanded, gDNA was extracted from the cell clones. The miR-155* TALEN pair-targeted region was amplified by PCR and subjected to TOPO cloning and sequencing. (B,C) Representative cell clones showing bi-allelic mutations (B) or mono-allelic mutations (C). In the sequence alignment graph, the left and right TALEN binding sites are highlighted in yellow and the miR-155* region is in the red box.