2. Academic Validation
  3. Nuclear Localization of Huntingtin mRNA Is Specific to Cells of Neuronal Origin

Nuclear Localization of Huntingtin mRNA Is Specific to Cells of Neuronal Origin

  • Cell Rep. 2018 Sep 4;24(10):2553-2560.e5. doi: 10.1016/j.celrep.2018.07.106.
Marie-Cécile Didiot 1,  Chantal M Ferguson 2,  Socheata Ly 2,  Andrew H Coles 2,  Abigail O Smith 2,  Alicia A Bicknell 2,  Lauren M Hall 2,  Ellen Sapp 3,  Dimas Echeverria 2,  Athma A Pai 2,  Marian DiFiglia 3,  Melissa J Moore 2,  Lawrence J Hayward 4,  Neil Aronin 5,  Anastasia Khvorova 6
Affiliations

Affiliations

  • 1 RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA. Electronic address: marie.didiot@umassmed.edu.
  • 2 RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • 3 MassGeneral Institute for Neurodegenerative Disease, Massachusetts General Hospital, Charlestown, MA 02129, USA.
  • 4 Department of Neurology, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • 5 RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA; Department of Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USA.
  • 6 RNA Therapeutics Institute, University of Massachusetts Medical School, Worcester, MA 01605, USA; Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA 01605, USA. Electronic address: anastasia.khvorova@umassmed.edu.
PMID: 30184490 DOI: 10.1016/j.celrep.2018.07.106
Abstract

Huntington's disease (HD) is a monogenic neurodegenerative disorder representing an ideal candidate for gene silencing with oligonucleotide therapeutics (i.e., Antisense Oligonucleotides [ASOs] and small interfering RNAs [siRNAs]). Using an ultra-sensitive branched fluorescence in situ hybridization (FISH) method, we show that ∼50% of wild-type HTT mRNA localizes to the nucleus and that its nuclear localization is observed only in neuronal cells. In mouse brain sections, we detect Htt mRNA predominantly in neurons, with a wide range of Htt foci observed per cell. We further show that siRNAs and ASOs efficiently eliminate cytoplasmic HTT mRNA and HTT protein, but only ASOs induce a partial but significant reduction of nuclear HTT mRNA. We speculate that, like other mRNAs, HTT mRNA subcellular localization might play a role in important neuronal regulatory mechanisms.

Keywords

ASOs; CAG repeat RNA foci; HTT mRNA; Huntington’s disease; RNA fluorescence in situ hybridization; confocal microscopy; siRNAs.

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