• Open Access

Selective silencing by RNAi of a dominant allele that causes amyotrophic lateral sclerosis

Authors

  • Hongliu Ding,

    1. Departments of Biochemistry and Molecular Pharmacology,
    Search for more papers by this author
    • *

      These authors contributed equally to the paper.

  • Dianne S. Schwarz,

    1. Departments of Biochemistry and Molecular Pharmacology,
    Search for more papers by this author
    • *

      These authors contributed equally to the paper.

  • Alex Keene,

    1. Departments of Biochemistry and Molecular Pharmacology,
    Search for more papers by this author
  • El Bachir Affar,

    1. Department of Pathology, Harvard Medical School, 200 Longwood Ave, Boston, MA 02115, USA
    Search for more papers by this author
  • Laura Fenton,

    1. Departments of Biochemistry and Molecular Pharmacology,
    Search for more papers by this author
  • Xugang Xia,

    1. Departments of Biochemistry and Molecular Pharmacology,
    Search for more papers by this author
  • Yang Shi,

    1. Department of Pathology, Harvard Medical School, 200 Longwood Ave, Boston, MA 02115, USA
    Search for more papers by this author
  • Phillip D. Zamore,

    Corresponding author
    1. Departments of Biochemistry and Molecular Pharmacology,
      Zuoshang Xu, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St, Worcester, MA 01605–2324, USA. Tel.: +1 508 856 3309; fax: +1 508 856 8390; e-mail: zuoshang.xu@umassmed.edu
      Phillip Zamore, as above. Tel.: +1 508 856 2191; fax: +1 508 856 2003; e-mail: phillip.zamore@umassmed.edu
    Search for more papers by this author
  • Zuoshang Xu

    Corresponding author
    1. Departments of Biochemistry and Molecular Pharmacology,
    2. Cell Biology, and
    3. Neuroscience Program, University of Massachusetts Medical School, 364 Plantation St, Worcester, MA 01605–2324, USA
      Zuoshang Xu, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St, Worcester, MA 01605–2324, USA. Tel.: +1 508 856 3309; fax: +1 508 856 8390; e-mail: zuoshang.xu@umassmed.edu
      Phillip Zamore, as above. Tel.: +1 508 856 2191; fax: +1 508 856 2003; e-mail: phillip.zamore@umassmed.edu
    Search for more papers by this author

Zuoshang Xu, Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364 Plantation St, Worcester, MA 01605–2324, USA. Tel.: +1 508 856 3309; fax: +1 508 856 8390; e-mail: zuoshang.xu@umassmed.edu
Phillip Zamore, as above. Tel.: +1 508 856 2191; fax: +1 508 856 2003; e-mail: phillip.zamore@umassmed.edu

Summary

RNA interference (RNAi) can achieve sequence-selective inactivation of gene expression in a wide variety of eukaryotes by introducing double-stranded RNA corresponding to the target gene. Here we explore the potential of RNAi as a therapy for amyotrophic lateral sclerosis (ALS) caused by mutations in the Cu, Zn superoxide dismutase (SOD1) gene. Although the mutant SOD1 is toxic, the wild-type SOD1 performs important functions. Therefore, the ideal therapeutic strategy should be to selectively inhibit the mutant, but not the wild-type SOD1 expression. Because most SOD1 mutations are single nucleotide changes, to selectively silence the mutant requires single-nucleotide specificity. By coupling rational design of small interfering RNAs (siRNAs) with their validation in RNAi reactions in vitro and in vivo, we have identified siRNA sequences with this specificity. A similarly designed sequence, when expressed as small hairpin RNA (shRNA) under the control of an RNA polymerase III (pol III) promoter, retains the single-nucleotide specificity. Thus, RNAi is a promising therapy for ALS and other disorders caused by dominant, gain-of-function gene mutations.

Ancillary