Targeting transfer rna for the suppression of nonsense mutations in messenger rna
AssigneeThe Regents of the University of California
InventorsPrashant Mali, Dhruva Katrekar, Thomas Morton
Life of this patent
Estimating the term…
Abstract
The disclosure provides for a targeting transfer RNA (ttRNA) that that suppresses nonsense mutations in messenger RNA, that comprises an anticodon sequence that binds to a stop codon and a variable loop sequence that comprises an RNA aptamer that has strong binding affinity to an RNA binding protein; and methods of use thereof.
Claims84
A targeting transfer RNA (ttRNA) comprising:
- a polynucleotide having a general structure from 5′ to 3′ of:
- 5′-(D-Loop domain)-(anticodon loop domain)-(variable loop domain)-(T loop domain)-3′,
- wherein the anticodon loop domain comprises a sequence selected from N1UCTAN2N3;
- N1UUUAN2N3;
- or N1UUCAN2N3, wherein N1 is a pyrimidine, N2 is a purine and N3 is any of U, A or C, that binds to a stop codon sequence of an mRNA,
- wherein the variable loop domain is at least about 12 to about 30 nucleotides in length and which comprises a sequence that is selectively bound by a protein or an agent tag, and
- wherein the ttRNA at least partially suppresses nonsense mutations in messenger RNA.
The ttRNA of any one of the preceding claims, wherein the polynucleotide is engineered from a tRNA for serine or arginine.
The ttRNA of any one of the preceding claims, wherein the ttRNA has a cloverleaf-like structure.
The ttRNA of any one of the preceding claims, wherein the D-Loop domain and T-Loop domain of the polynucleotide comprises a naturally occurring sequence from a human tRNA.
The ttRNA of any one of the preceding claims, wherein the anticodon loop domain binds to a stop codon having a DNA sequence of ‘TAG’ or an RNA sequence of ‘UAG.’
The ttRNA of any one of the preceding claims, the variable loop domain comprises a sequence that is selectively bound by an RNA binding protein.
The ttRNA of any one of the preceding claims, where the variable loop domain comprises a sequence for BoxBv1.
The ttRNA of any one of the preceding claims, where the polynucleotide comprises a sequence with at least 70% sequence identity to a sequence selected from (1) to (8):
- (1) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- GGCCCTGAAAAAGGGCCGCGCAGGTTCGAATCCTGCCGACTACG;
- (2) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- CGGCCCTGAAAAAGGGCCGCGCAGGTTCGAATCCTGCCGACTACG;
- (3) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- GGGGACATGAGGATCACCCATGTCCCGCGCAGGTTCGAATCCTGCCGA
- CTACG;
- (4) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- AGCTTATCCATTGCACTCCGGATAAGCTGCGCAGGTTCGAATCCTGCC
- GACTACG;
- (5) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- GGCCCAGTGGTCTTTGTAGACTGCCTGATGGCCGCGCAGGTTCGAATC
- CTGCCGACTACG;
- (6) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- GGCCAGATCTGAGCCTGGGAGCTCTCTGGCCGCGCAGGTTCGAATCCT
- GCCGACTACG;
- (7) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- CCAAAGGCTCTTCTCAGAGCCACCCAGCGCAGGTTCGAATCCTGCCGA
- CTACG;
- and
- (8) GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATT
- GGCTCTTCTCAGAGCCGCGCAGGTTCGAATCCTGCCGACTACG.
The ttRNA of any one of the preceding claims, wherein the polynucleotide comprises the sequence of:
- GTAGTCGTGGCCGAGTGGTTAAGGCGATGGACTCTAAATCCATTGGCC
- CTGAAAAAGGGCCGCGCAGGTTCGAATCCTGCCGACTACG.
A vector that encodes the ttRNA of any one of the preceding claims.
An engineered targeting transfer RNA (ttRNA) comprising a modified variable loop, wherein the modified variable loop comprises a hairpin loop that binds to an RNA binding protein to a greater extent than a tRNA comprising an unmodified variable loop
Dependencies are inferred from the claim text.