Generation of an isogenic, gene-corrected control cell line of the spinocerebellar ataxia type 2 patient-derived iPSC line H196

Adele G. Marthaler, Benjamin Schmid, Alisa Tubsuwan, Ulla B. Poulsenc, Alexander F. Engelbrecht,Ulrike A. Mau-Holzmann , Poul Hyttel , Jørgen E. Nielsen, Troels T. Nielsenb, Bjørn Holst
Source: Stem Cell Res
Publication Date: (2016)
Issue: 16: 162-165
Research Area:
Stem Cells
Cells used in publication:
Induced Pluripotent Stem Cell (iPS), human
Species: human
Tissue Origin:
4D-Nucleofector® X-Unit
patient derived iPSC were growing on matrigel coated dishes and detached with accutase. 1x10e6 cells were co-nucleofected with 2µg of each CRISPR/Cas9 plasmid Addgene plasmid ID 48139) + 1µg of the restistance marker plasmid program CA-167 P3 primary cell solution cells were plated on matrigel coated dishes in E8 medium supplemented with ROCKinhibitor; 24h post nucleofection antibiotic selection was started.
Spinocerebellar ataxia type 2 (SCA2) is a neurodegenerative disease primarily affecting the cerebellum. Very little is known about the molecular mechanisms underlying the disease and, to date, no cure or treatment is available. We have successfully generated bona fide induced pluripotent stem cell (iPSC) lines of SCA2 patients in order to study a disease-specific phenotype. Here, we demonstrate the gene correction of the iPSC line H196 clone 7 where we have exchanged the expanded CAG repeat of the ATXN2 gene with the normal length found in healthy alleles. This gene corrected cell line will provide the ideal control to model SCA2 by iPSC technology.