Huntington’s disease

Isogenic iPSC lines - Huntington's disease

Isogenic HD model
Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the HTT gene. Here we generated isogenic iPSC-based HD model by introducing 69 CAG triplets into HTT through CRISPR/Cas9-mediated homologous recombination. The obtained isogenic cell system can serve as a valuable model for the investigation into various parameters of HD and for screening of potential drugs without worries about background-related variability.

Isogenic iPSC lines - Huntington's disease
(A) The scheme of homologous recombination between wild-type HTT and the donor construct. HTT-F and HTT-R: primers for the detection of the insertion of the expanded CAG repeat tract into HTT; (B) PCR analysis of HTT allele lengths in the mutant cell clones; (C) Western blot analysis of HTT expression in the mutant cells; (D) Fragment analysis of PCR products by capillary electrophoresis. Green squares and arrows indicate the number of CAG repeats in the HTT alleles; (E) Immunofluorescence staining of isogenic HD iPSCs with antibodies against transcription factors OCT3/4, NANOG, and SOX2 and surface antigen TRA-1-60; (F) Immunofluorescence staining confirming the expression of markers of three germ layers’ derivatives after spontaneous differentiation of isogenic HD iPSCs. Scale bars: 100 µm.

Isogenic HD model
Huntington’s disease (HD) is a severe neurodegenerative disorder caused by a CAG triplet expansion in the first exon of the HTT gene. Here we generated isogenic iPSC-based HD model by introducing 69 CAG triplets into HTT through CRISPR/Cas9-mediated homologous recombination. The obtained isogenic cell system can serve as a valuable model for the investigation into various parameters of HD and for screening of potential drugs without worries about background-related variability.