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Updated: Dec 26, 2025
Generation of Defined Genomic Modifications Using CRISPR-CAS9 in Human Pluripotent Stem Cells
Published on: September 25, 2019
Generation of New Isogenic Models of Huntington's Disease Using CRISPR-Cas9 Technology
Magdalena Dabrowska1, Agata Ciolak2, Emilia Kozlowska2
1Department of Genome Engineering, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12/14, 61-704 Poznan, Poland.
Insights
Researchers created Huntington's disease (HD) cell models using genome engineering. These isogenic cell lines aid in understanding HD pathogenesis and testing new therapeutic reagents for the fatal neurodegenerative disorder.
Area of Science:
- Neuroscience
- Genetics
- Molecular Biology
Background:
- Huntington's disease (HD) is a fatal, monogenic neurodegenerative disorder.
- The exact pathogenesis of HD remains unclear, and effective therapies are lacking.
- Genome engineering offers new avenues for disease modeling and drug discovery.
Purpose of the Study:
- To generate and validate isogenic cell models for Huntington's disease research.
- To assess the utility of these models for testing therapeutic interventions.
- To provide a comprehensive set of tools for investigating HD.
Main Methods:
- Generation of homozygous HEK 293T cell lines with varying CAG repeat lengths in the huntingtin gene (HTT).
- Application of the CRISPR-Cas9 system to correct the HD mutation in human induced pluripotent stem cells.
- Creation of an HTT gene knockout using CRISPR-Cas9.
Main Results:
- Successfully generated a series of homozygous HEK 293T cell lines with defined CAG repeat expansions.
- Demonstrated the utility of these cell lines for screening therapeutic reagents.
- Developed isogenic human induced pluripotent stem cell lines with corrected and knocked-out HTT genes.
Conclusions:
- The generated isogenic cell lines are valuable tools for studying Huntington's disease pathogenesis.
- These models facilitate the testing of potential therapeutic agents for HD.
- A comprehensive suite of isogenic cell lines is now available for HD research.
Abstract:
Huntington's disease (HD) is a fatal neurodegenerative disorder caused by the expansion of CAG repeats in exon 1 of the huntingtin gene (HTT). Despite its monogenic nature, HD pathogenesis is still not fully understood, and no effective therapy is available to patients. The development of new techniques such as genome engineering has generated new opportunities in the field of disease modeling and enabled the generation of isogenic models with the same genetic background. These models are very valuable for studying the pathogenesis of a disease and for drug screening. Here, we report the generation of a series of homozygous HEK 293T cell lines with different numbers of CAG repeats at the HTT locus and demonstrate their usefulness for testing therapeutic reagents. In addition, using the CRISPR-Cas9 system, we corrected the mutation in HD human induced pluripotent stem cells and generated a knock-out of the HTT gene, thus providing a comprehensive set of isogenic cell lines for HD investigation.
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