Compositions and Methods of Generating Novel amiRNA
AssigneeMirimus, Inc.
InventorsGiuseppe Militello, Christof Fellmann, Johannes Zuber
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Abstract
Provided herein are Compositions and Methods of Generating Novel amiRNA's. Provided herein are methods for producing or generating one or more amiRNAs; also provided are constructs and compositions useful in the methods. The methods and constructs provided in this disclosure are highly efficient methods for production of a new generation of amiRNAs.
Claims11
A method of generating amiRNAs and modifying a selected primary miRNA, comprising:
- Applying a base modifying step to produce a single stranded region of at least 9 nucleotides and a GU dinucleotide in position −13 and −14, respectively from a Drosha cleavage site to form a base modified construct;
- Applying a basal stem step to produce a double stranded RNA with a CHC motif and a bulge in position +8 from the Drosha cleavage site to form a base stem modified construct;
- Applying a loop modifying step to substitute the endogenous loop region of the primary miRNA with the loop of a miRE, or another loop with same number of nucleotides and the GUG motif in the same position, to form a loop modified construct;
- and
- Operably linking the base modified construct, the basal stem modified construct, and the loop modified construct with the primary miRNA into an amiRNA construct.
A heterologous or synthetic or artificial single-stranded ribonucleic acid (RNA) sequence comprising (i) a selected primary miRNA and the complement thereof, and (ii) a base region in between the selected primary miRNA and the complement thereof, wherein the nucleotide sequence of the base region is at least 75% sequence identity to SEQ ID NO:
- 1.
A heterologous or synthetic or artificial single-stranded ribonucleic acid (RNA) sequence comprising (i) a selected primary miRNA and a complement thereof, and (ii) a base region and a basal stem in between the selected primary miRNA and a loop region, wherein the nucleotide sequence of the base region, the base stem region, and the loop region is at least 75% sequence identity to SEQ ID NOs:
- 3-9.
Dependencies are inferred from the claim text.