Oligomers
Molecules are provided for inducing or facilitating exon skipping in forming spliced mRNA products from pre-mRNA molecules in cells. The molecules may be provided directly as oligonucleotides or expression products of vectors that are administered to a subject. High rates of skipping can be achieved. High rates of skipping reduce the severity of a disease like Duchene Muscular Dystrophy so that the disease is more like Becker Muscular Dystrophy. This is a severe reduction in symptom severity and mortality.
1. An oligomer for ameliorating Duchenne muscular dystrophy (DMD), the oligomer comprising at least 25 contiguous bases of:
a)
(SEQ ID NO: 10)
CXG XXG CCX CCG GXX CXG AAG GXG XXC XXG;
or
b)
(SEQ ID NO: 12)
XXG CCX CCG GXX CXG AAG GXG XXC XXG XAC;
wherein X=U or T, wherein the sequence of the at least 25 contiguous bases can vary from SEQ ID NO: 10 or 12 at up to two base positions, and wherein the oligomer comprises one or more synthetically modified nucleotides.
2. The oligomer according to claim 1 , wherein the oligomer can bind to a target site to cause exon skipping of exon 53 of the dystrophin gene.
3. The oligomer according to claim 1 , wherein the oligomer can bind to a target site to cause exon skipping of an exon of the dystrophin gene at a rate of at least 50%.
4. The oligomer according to claim 1 , wherein the oligomer can bind to a target site to cause exon skipping of an exon of the dystrophin gene at a rate of at least 70%.
5. The oligomer according to claim 1 , wherein the oligomer can bind to a target site to cause exon skipping of an exon of the dystrophin gene at a rate of at least 90%.
6. The oligomer according to claim 1 , wherein the one or more synthetically modified nucleotides are modified at the 2′ position of the ribose.
7. The oligomer according to claim 6 , wherein the one or more synthetically modified nucleotides are 2′-O-methyl oligonucleotide.
8. The oligomer according to claim 1 , wherein the one or more synthetically modified nucleotides are a phosphorodiamidate morpholino oligonucleotide.
9. The oligomer according to claim 1 , wherein the oligomer is at least 28 and at most 32 bases in length.
10. The oligomer according to claim 1 , wherein the oligomer is at least 29 and at most 31 bases in length.
11. The oligomer according to claim 1 , wherein the oligomer is conjugated to or complexed with a targeting protein that targets the oligomer to muscle tissue.
12. The oligomer according to claim 1 , wherein the oligomer is conjugated to or complexed with a drug for treating Duchenne muscular dystrophy.
13. The oligomer according to claim 1 , wherein the oligomer is conjugated to or complexed with an arginine-rich cell penetrating peptide.
14. A pharmaceutical composition for ameliorating DMD, the composition comprising an oligomer according to claim 1 , and a pharmaceutically acceptable carrier, adjuvant or vehicle.