TAL-Based Editing of iPSCs for the Study of Parkinson's Disease

Due to the inherent biological variability between cell lines of different backgrounds, the study of subtle phenotypic changes associated with disease states when using iPSC derived materials requires isogenic control lines for proper comparison-- that is, cell lines that differ in a defined location but that are otherwise identical at the genetic level.  This presentation will describe the approaches we've taken to modifying genes involved in Parkinson's Disease and related disorders.  For example, modifying the glucocerebrosidase gene in induced pluripotent stem cells (iPSCs) using TAL-based engineering technology presented several challenges, including the presence of an adjacent and highly homologous pseudogene.  Also, creation of both single- and double-knockouts of the alpha-synuclein gene in iPSCs also presented unique challenges that influenced the approach taken in order to be successful.  Both of these examples will describe the unique problems we encountered in the project, as well as the strategies taken to overcome these challenges.  Initial phenotypic comparisons between the patient derived, TAL-edited lines that we have generated will be presented.