Reversal of Phenotypic Abnormalities by CRISPR/Cas9-Mediated Gene Correction in Huntington Disease Patient-Derived Induced Pluripotent Stem Cells

• DOI: 10.1016/j.stemcr.2017.01.022
• Publication Date: 2017-02-23T19:17:29Z
• Authors: Xiaohong Xu; Yilin Tay; Bernice Sim; Su-In Yoon; Yihui Huang; Jolene Ooi; Kagistia Hana Utami; Amin Ziaei; Bryan Ng; Carola Radulescu; Donovan Low; Alvin Yu Jin Ng; Marie Loh; Byrappa Venkatesh; Florent Ginhoux; George J. Augustine; Mahmoud A. Pouladi
• Keywords: 0301 basic medicine; Medicine (General); hiPSC; transcriptional dysrgulation; disease modeling; genetic editing; Developmental; Biology (General); Cell Self Renewal; CAS9; TRANSGENIC MICE; Gene Editing; Neurons; human induced pluripotent stem cell (hiPSC); Gene Expression Regulation, Developmental; Cell Differentiation; DEFECTS; Huntington disease; Mitochondria; 3. Good health; DNA-Binding Proteins; DIFFERENTIATION; Huntington Disease; Phenotype; neurodegenerative disorders; Gene Targeting; Life Sciences & Biomedicine; 570; CHCHD2; QH301-705.5; MODELS; Induced Pluripotent Stem Cells; INHIBITION; 610; Article; Cell Line; Mitochondrial Proteins; 03 medical and health sciences; R5-920; Cell & Tissue Engineering; mitochondrial dysfunction; Humans; STRIATAL PROJECTION NEURONS; Science & Technology; MUTATIONS; Cell Biology; Electrophysiological Phenomena; Gene Expression Regulation; CRISPR-Cas Systems; disease phenotypes; Transcription Factors

Highlights•A CRISPR-Cas9 and PiggyBac-based approach allows efficient correction of HD hiPSCs•The corrected hiPSCs can be differentiated into synaptically active neurons•Correction gene mutation reverses a number phenotypic abnormalities•Isogenic help distinguish from genetic background-related effectsSummaryHuntington disease (HD) is dominant neurodegenerative disorder caused by CAG repeat expansion in HTT. Here we report human induced pluripotent stem cells (hiPSCs) using piggyBac transposon-based approach. We show that both isogenic excitable, forebrain neurons. further demonstrate abnormalities hiPSC-derived neural cells, including impaired rosette formation, increased susceptibility to growth factor withdrawal, deficits mitochondrial respiration, are rescued controls. Importantly, genome-wide expression analysis, apparent differences detected between non-related healthy control lines absent lines, suggesting these likely related background rather than HD-specific effects. Our study demonstrates associated abnormalities, the importance controls for modeling hiPSCs.Graphical abstract