Vivo-Morpholinos: a non-peptide transporter delivers Morpholinos into a wide array of mouse tissues

Biotechniques. 2008 Dec;45(6):613-4, 616, 618 passim. doi: 10.2144/000113005.

Abstract

We have developed a new transporter structure that provides effective delivery of Morpholino antisense oligomers into a wide variety of tissues in living mice. This transporter comprises a dendritic structure assembled around a triazine core which serves to position eight guanidinium head groups in a conformation effective to penetrate cell membranes. This transporter structure is conjugated to a Morpholino oligomer to form a delivery-enabled product referred to as a Vivo-Morpholino. Vivo-Morpholinos are shown to effectively enter and function within cultured cells in the presence of 100% serum using a rigorous positive test system based on correction of a defined splicing error in a pre-messenger RNA. In addition, Vivo-Morpholinos are demonstrated to enter into a wide variety of tissues in a similar positive test system in transgenic mice, as evidenced by correction of the targeted splicing error in all tissues assessed, including near-complete splice correction in the small intestine, colon, stomach, liver kidney, and a number of muscles. Finally, Vivo-Morpholinos, which target the exon-skipping of exon 23 harboring a premature termination codon in the mdx mouse model, effectively restore the reading frame of dystrophin and restore expression of a functional dystrophin protein.

MeSH terms

  • Animals
  • Cells, Cultured
  • Disease Models, Animal
  • Drug Delivery Systems / methods*
  • Dystrophin / genetics
  • Dystrophin / metabolism
  • Fluorescent Antibody Technique
  • Gene Expression
  • Green Fluorescent Proteins / metabolism
  • Guanidine / metabolism
  • Mice
  • Mice, Inbred mdx
  • Mice, Transgenic
  • Muscular Dystrophy, Duchenne
  • Mutation
  • Oligonucleotides, Antisense / metabolism*
  • Peptides / metabolism
  • RNA Splicing
  • Reverse Transcriptase Polymerase Chain Reaction
  • Targeted Gene Repair / methods*
  • Triazines / metabolism

Substances

  • Dystrophin
  • Endo-Porter
  • Oligonucleotides, Antisense
  • Peptides
  • Triazines
  • Green Fluorescent Proteins
  • Guanidine