Analysis of Gene Targeting Techniques for Huntington’s Disease and Gene Expression in Human Cells

Gemstone Team CHANGEHuntington’s disease (HD) is an inherited neurodegenerative disorder that is caused by a CAG trinucleotide repeat expansion in the huntingtin (HTT) gene. Our team performed a literature analysis to investigate the current state of research for treating HD and identified a new technology called prime editing that could be applied to HD in combination with single nucleotide polymorphisms (SNPs). We found that at least 729 SNPs within the HTT gene are compatible with our proposed approach. Experimentally, we performed preliminary studies using Western Blots and RT-qPCR to examine the differences in expression of HTT in a variety of cell lines. Our literature-based work suggests that prime editing is a promising tool for addressing the basis of a variety of genetic disorders. Our experimental-based work confirms that human fibroblast cells express HTT and therefore may be used in proof of concept studies of gene targeting techniques to address HD

Gene Targeting Techniques for Huntington's Disease

Huntington’s disease (HD) is an autosomal neurodegenerative disorder caused by extended trinucleotide CAG repetition in the HTT gene. Although this mutation in the HTT gene is mostly associated with neurological and physical symptoms that HD typically exhibits, wild-type Huntingtin protein (HTT) is involved in a variety of cellular functions such as vesicle transportation, cell division, transcription regulation, autophagy, and tissue maintenance. The main cause of HD symptoms is due to aggregation and accumulation of mutant HTT (mHTT) proteins in neurons. In this review, we discuss multiple approaches targeting DNA and RNA to reduce mHTT expression. These approaches are categorized into non-allele-specific silencing and allele-specific-silencing using SNPs and haplogroup analysis, and the possible limitations of targeting mHTT is also discussed. Additionally, this review discusses am potential appliction of recent CRISPR prime editing technology in treating HD