Allele-Specific Reduction of the Mutant Huntingtin Allele Using Transcription Activator-Like Effectors in Human Huntington's Disease Fibroblasts

Cell Transplantation

Article

Teresa Tempkin¹  Alexandra Duffy¹  Vicki Wheelock¹  David J. Segal²  Josh Gutierrez³  Whitney Cary³  Geralyn Annett³  Kyle D. Fink³  Audrey Torrest³  William Gruenloh³  Joseph S. Anderson³  Johnathon D. Anderson³  Jan A. Nolta³  Stefanos Kalomoiris³  Peter Deng⁴  Anvita Komarla⁴ 

[1] Department of Neurology, University of California Davis Health Systems, Sacramento, CA, USA;Genome Center, MIND Institute, and Biochemistry and Molecular Medicine, University of California, Davis, CA, USA;Stem Cell Program and Institute for Regenerative Cures, University of California Davis Health Systems, Sacramento, CA, USA;Stem Cell Program and Institute for Regenerative Cures, University of California Davis Health Systems, Sacramento, CA, USA;Genome Center, MIND Institute, and Biochemistry and Molecular Medicine, University of California, Davis, CA, USA;

关键词: Huntington's disease (HD);    Transcription activator-like effector (TALE);    Gene therapy;    Allele-specific silencing;   

DOI  :  10.3727/096368916X690863

 received in 2015-10-31, accepted in 2016-01-28,  发布年份 2016

来源: Sage Journals

【 摘 要 】

Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by an abnormal expansion of CAG repeats. Although pathogenesis has been attributed to this polyglutamine expansion, the underlying mechanisms through which the huntingtin protein functions have yet to be elucidated. It has been suggested that postnatal reduction of mutant huntingtin through protein interference or conditional gene knockout could prove to be an effective therapy for patients suffering from HD. For allele-specific targeting, transcription activator-like effectors (TALE) were designed to target single-nucleotide polymorphisms (SNP) in the mutant allele and packaged into a vector backbone containing KRAB to promote transcriptional repression of the disease-associated allele. Additional TALEs were packaged into a vector backbone containing heterodimeric FokI and were designed to be used as nucleases (TALEN) to cause a CAG-collapse in the mutant allele. Human HD fibroblasts were treated with each TALE-SNP or TALEN. Allele-expression was measured using a SNP-genotyping assay and mutant protein aggregation was quantified with Western blots for anti-ubiquitin. The TALE-SNP and TALEN significantly reduced mutant allele expression (p < 0.05) when compared to control transfections while not affecting expression of the nondisease allele. This study demonstrates the potential of allele-specific gene modification using TALE proteins, and provides a foundation for targeted treatment for individuals suffering from Huntington's or other genetically linked diseases.

【 授权许可】

Unknown   
© 2016 Cognizant, LLC.

【 图 表 】

Figure 1