Allele-specific silencing of mutant huntingtin in rodent brain and human stem cells

PLoS One. 2014 Jun 13;9(6):e99341. doi: 10.1371/journal.pone.0099341. eCollection 2014.


Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder resulting from polyglutamine expansion in the huntingtin (HTT) protein and for which there is no cure. Although suppression of both wild type and mutant HTT expression by RNA interference is a promising therapeutic strategy, a selective silencing of mutant HTT represents the safest approach preserving WT HTT expression and functions. We developed small hairpin RNAs (shRNAs) targeting single nucleotide polymorphisms (SNP) present in the HTT gene to selectively target the disease HTT isoform. Most of these shRNAs silenced, efficiently and selectively, mutant HTT in vitro. Lentiviral-mediated infection with the shRNAs led to selective degradation of mutant HTT mRNA and prevented the apparition of neuropathology in HD rat's striatum expressing mutant HTT containing the various SNPs. In transgenic BACHD mice, the mutant HTT allele was also silenced by this approach, further demonstrating the potential for allele-specific silencing. Finally, the allele-specific silencing of mutant HTT in human embryonic stem cells was accompanied by functional recovery of the vesicular transport of BDNF along microtubules. These findings provide evidence of the therapeutic potential of allele-specific RNA interference for HD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Brain / cytology*
  • Brain / metabolism
  • Cells, Cultured
  • Disease Models, Animal
  • Embryonic Stem Cells / cytology
  • Genetic Therapy / methods*
  • HEK293 Cells
  • Humans
  • Huntingtin Protein
  • Huntington Disease / genetics
  • Huntington Disease / therapy*
  • In Vitro Techniques
  • Male
  • Mice
  • Mutant Proteins / antagonists & inhibitors*
  • Mutant Proteins / genetics
  • Nerve Tissue Proteins / genetics*
  • Polymorphism, Single Nucleotide
  • RNA Isoforms / metabolism
  • RNA Stability
  • RNA, Small Interfering / genetics*
  • Rats
  • Rats, Wistar


  • HTT protein, human
  • Huntingtin Protein
  • Mutant Proteins
  • Nerve Tissue Proteins
  • RNA Isoforms
  • RNA, Small Interfering

Grants and funding

This work was supported by the Agence Nationale de la Recherche? [ANR-2006-MRAR-043-01], FP6-NeuroNe [LSHM-CT-2004-512039], FP6-Clinigene [LSH-2004-1.2.4-3], the Atomic Energy Commission, FP6 STEM-HD, DIM-STEM POLE Region Ile-de-France fellowship to [F.B-R] and the Swiss National Science Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.