Vivo-Morpholinos: a non-peptide transporter delivers Morpholinos into a wide array of mouse tissues.
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.
Morcos
Paul A
PA
Gene Tools, LLC, Philomath, OR 97370, USA. pmorcos@gene-tools.com
Li
Yongfu
Y
Jiang
Shan
S
D000818:Animals; D002478:Cells, Cultured; D004195:Disease Models, Animal; D016503:Drug Delivery Systems / Q000379:methods*; D016189:Dystrophin / Q000235:genetics / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000378:metabolism; D019791:Guanidine / Q000378:metabolism; D051379:Mice; D018101:Mice, Inbred mdx; D008822:Mice, Transgenic; D020388:Muscular Dystrophy, Duchenne; D009154:Mutation; D016376:Oligonucleotides, Antisense / Q000378:metabolism*; D010455:Peptides / Q000378:metabolism; D012326:RNA Splicing; D020133:Reverse Transcriptase Polymerase Chain Reaction; D052416:Targeted Gene Repair / Q000379:methods*; D014227:Triazines / Q000378:metabolism
D016189:Dystrophin; C511217:Endo-Porter; D016376:Oligonucleotides, Antisense; D010455:Peptides; D014227:Triazines; D049452:Green Fluorescent Proteins; D019791:Guanidine