Trans-excision-splicing ribozyme and methods of use
AssigneeUniversity of Kentucky Research Foundation
InventorsStephen M. Testa, Michael A. Bell
Life of this patent
Estimating the term…
Abstract
A group I intron-derived ribozyme which binds RNA in trans, excises an internal segment from within the RNA, and splices the remaining 5′ and 3′ ends of the RNA back together (the trans-excision-splicing reaction) is disclosed. The excised segment can be as long as 28 nucleotides, or more, and as little as one nucleotide. The ribozymes of the invention are easily modified to alter their sequence specificity. Such ribozymes represent a new and potentially powerful class of generally adaptable genetic therapeutics.
Claims19
1-9. (canceled)
A method of removing a non-native nucleotide sequence from a target nucleic acid sequence comprising contacting the target nucleic acid sequence with a modified trans-excision-splicing Group I ribozyme comprising at least two modifiable recognition elements, wherein at least one of said recognition elements is complementary to the non-native target sequence and at least one of said recognition elements stabilizes binding of the ribozyme to a trans-excision splicing (TES) reaction intermediate product, and wherein the ribozyme catalyzes a specific excision of the non-native target sequence and splices together the 5′ and 3′ ends of the substrate created by the excision.
A method of treating a disease associated with a genetic mutation comprising administering to a patient in need thereof a modified trans-excision-splicing Group I ribozyme comprising at least two modifiable recognition elements, wherein at least one of said recognition elements is complementary to non-native target sequence associated with the disease and at least one of said recognition elements stabilizes binding of the ribozyme to a trans-excision splicing (TES) reaction intermediate product, and wherein the ribozyme catalyzes a specific excision of the non-native target sequence and splices together the 5′ and 3′ ends of the substrate created by the excision.
(canceled)
A method of removing an internal expanded triplet repeat sequence from an RNA molecule comprising contacting the RNA molecule with a modified trans-excision-splicing Group I ribozyme comprising at least two modifiable recognition elements, wherein at least one of said recognition elements is complementary to non-native target RNA sequence within a substrate and at least one of said recognition elements stabilizes binding of the ribozyme to a trans-excision splicing (TES) reaction intermediate product, and wherein the ribozyme catalyzes a specific excision of the non-native target RNA sequence and splices the 5′ end of the substrate created by the excision to an ωG of the 3′ end of the substrate created by the excision.
A method of removing a premature stop codon from a mutant RNA sequence comprising contacting the mutant RNA sequence with a modified trans-excision-splicing Group I ribozyme comprising at least two modifiable recognition elements wherein at least one of said recognition elements is complementary to non-native target RNA sequence within a substrate and at least one of said recognition elements stabilizes binding of the ribozyme to a trans-excision splicing (TES) reaction intermediate product and wherein the ribozyme catalyzes a specific excision of the non-native target RNA sequence and splices the 5′ end of the substrate created by the excision to an ωG of the 3′ end of the substrate created by the excision.
Dependencies are inferred from the claim text.