Use of vivo-morpholinos for control of protein expression in the adult rat brain

J Neurosci Methods. 2012 Jan 30;203(2):354-60. doi: 10.1016/j.jneumeth.2011.10.009. Epub 2011 Oct 17.

Abstract

Vivo-morpholinos are commercially available morpholino oligomers with a terminal octa-guanidinium dendrimer for enhanced cell-permeability. Existing evidence from systemically delivered vivo-morpholinos indicate that genetic suppression can last from days to weeks without evidence of cellular toxicity. However, intravenously delivered vivo-morpholinos are ineffective at protein suppression in the brain, and no evidence is available regarding whether intracranially delivered vivo-morpholinos effectively reduce target protein levels, or do so without inducing neurotoxicity. Here we report examples in which in vivo microinjection of antisense vivo-morpholinos directed against three different targets (xCT, GLT1, orexin) in two different brain regions resulted in significant suppression of protein expression without neurotoxicity. Expression was significantly suppressed at six to seven days post-administration, but returned to baseline levels within fourteen days. These results indicate that direct intracranial administration of vivo-morpholinos provides an effective means by which to suppress protein expression in the brain for one to two weeks.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antisense Elements (Genetics) / administration & dosage*
  • Antisense Elements (Genetics) / chemistry
  • Antisense Elements (Genetics) / genetics
  • Brain / cytology
  • Brain / drug effects
  • Brain / metabolism*
  • Male
  • Microinjections / methods
  • Morpholinos / administration & dosage*
  • Morpholinos / chemistry
  • Morpholinos / genetics
  • Nerve Tissue Proteins / antagonists & inhibitors
  • Nerve Tissue Proteins / biosynthesis*
  • Nerve Tissue Proteins / genetics*
  • Proteomics / methods*
  • RNA, Messenger / antagonists & inhibitors
  • RNA, Messenger / metabolism
  • Rats
  • Rats, Sprague-Dawley

Substances

  • Antisense Elements (Genetics)
  • Morpholinos
  • Nerve Tissue Proteins
  • RNA, Messenger