In vivo identification of therapeutic constructs from pooled candidates in HD model mice

J Huntingtons Dis. 2013;2(4):437-41. doi: 10.3233/JHD-130073.

Abstract

Rapidly identifying targets for Huntington's Disease (HD) therapeutics in relevant mouse models could hasten the development of patient interventions. We have recently described a method for rapidly and quantitatively measuring the progression of HD-like symptoms in mouse models. Because this method uses flow cytometry to measure GFP levels in affected neurons, it is amenable to pooled approaches. Here we describe a continuation of this work, using pools of shRNA-delivering AAV vectors and high throughput sequencing to determine which hairpins in a mixed population are most effective at preventing the transcriptional dysregulation phenotype of R6/2 mice.

Keywords: Huntington's Disease; Neurodegeneration; gene expression; neuroprotection; pooled screening; viral vectors.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Dependovirus
  • Disease Models, Animal
  • Flow Cytometry
  • Genetic Therapy / methods*
  • Genetic Vectors / metabolism
  • Green Fluorescent Proteins / genetics
  • Huntington Disease / genetics*
  • Huntington Disease / metabolism
  • Huntington Disease / pathology
  • Mice
  • Mice, Transgenic
  • Mutant Proteins / genetics
  • Neurons / metabolism
  • Neurons / pathology
  • Serotonin Plasma Membrane Transport Proteins / genetics

Substances

  • Mutant Proteins
  • Serotonin Plasma Membrane Transport Proteins
  • Slc6a4 protein, mouse
  • Green Fluorescent Proteins