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US 11,453,876 B2Granted
Compositions and methods for identifying polynucleotides of interest
AssigneeThe Regents of the University of California
InventorsPaul A. Feldstein, James E. Lincoln
Life of this patent
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Abstract
Provided are constructs and methods useful for the screening and identification of polynucleotide sequences of interest, in particular promoters, RNA stability modifying sequences and transcriptional modifying sequences.
Claims20
A DNA plasmid comprising operably linked polynucleotides encoding:
- (a) a mini-monomer cassette, comprising a first ribozyme cleavage site and a second ribozyme cleavage site, and comprising between the first ribozyme cleavage site and the second ribozyme cleavage site:
- (i) a ribozyme catalytic core;
- (ii) a multiple cloning site polynucleotide comprising plurality of restriction endonuclease cleavage sites;
- and
- (iii) a reverse primer annealing polynucleotide located upstream or 5′ to a forward primer annealing polynucleotide;
- and
- (b) a plasmid backbone, comprising an origin of replication.
4 more elements
A DNA plasmid comprising operably linked polynucleotides encoding:
- (a) a mini-monomer cassette, comprising a first ribozyme cleavage site and a second ribozyme cleavage site, and comprising between the first ribozyme cleavage site and the second ribozyme cleavage site:
- (i) a ribozyme catalytic core;
- (ii) a reverse primer annealing polynucleotide;
- (iii) a forward primer annealing polynucleotide;
- and
- (iv) an inserted polynucleotide suspected of comprising a promoter;
- and
- (b) a plasmid backbone, comprising an origin of replication.
5 more elements
A DNA plasmid comprising operably linked polynucleotides encoding:
- (a) a test promoter, wherein the test promoter is functional in a eukaryotic cell or in a prokaryotic cell;
- (b) a mini-monomer cassette, comprising:
- (i) a first ribozyme cleavage site;
- (ii) a ribozyme catalytic core;
- (iii) a reverse primer annealing polynucleotide;
- (iv) a forward primer annealing polynucleotide;
- (v) an inserted polynucleotide suspected of comprising an RNA stability modifier or a transcription modifier;
- (vi) a second ribozyme cleavage site;
- (c) a plasmid backbone, comprising an origin of replication.
6 more elements
A DNA plasmid comprising operably linked polynucleotides encoding:
- (a) a test promoter, wherein the test promoter is functional in a eukaryotic cell or in a prokaryotic cell;
- (b) a first mini-monomer cassette, comprising:
- (i) a first ribozyme cleavage site;
- (ii) a first ribozyme catalytic core;
- (iii) a first reverse primer annealing polynucleotide;
- (iv) a first forward primer annealing polynucleotide;
- (v) a barcode polynucleotide
- (vi) a second ribozyme cleavage site;
- (c) a second mini-monomer cassette, comprising:
- (i) the second ribozyme cleavage site;
- (ii) a second ribozyme catalytic core;
- (iii) a second reverse primer annealing polynucleotide;
- (iv) a second forward primer annealing polynucleotide;
- (v) an inserted polynucleotide suspected of comprising an RNA stability modifier or a transcription modifier;
- (vi) a third ribozyme cleavage site;
- (d) a plasmid backbone, comprising an origin of replication.
13 more elements
Dependencies are inferred from the claim text.