System for regulating gene expression
AssigneeBaylor College of Medicine
InventorsLaising Yen, Liming Luo, Jocelyn Duen-Ya Jea
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Abstract
Compositions and methods relating to regulation of gene expression are described. In some embodiments, the present disclosure provides compositions and methods for the regulation of gene expression using nucleic acid constructs. In some embodiments, the present disclosure recognizes the utility of alternative splicing in regulation of gene expression in a nucleic acid construct. In some embodiments, the present disclosure recognizes the utility of regulating gene expression utilizing ligand-binding aptamers.
Claims29
A system for modulating gene expression, comprising a polyA aptamer polynucleotide,
- that comprises in a 5′ to 3′ direction:
- a) a 5′ splice donor site;
- b) an engineered intron with a
- a first 3′ splice acceptor site on the 3′ side of the engineered intron;
- and,
- a polyA switch comprising three ligand-binding aptamers each with one or more ligand binding pockets, a three way junction, and at least one polyA cleavage signal therein;
- wherein the first 3′ splice acceptor site is either located 5′ of the polyA switch or in the polyA switch;
- c) a second 3′ splice acceptor site;
- and
- d) a nucleic acid sequence encoding an expressible polypeptide;
- wherein absence of a ligand allows splicing between the 5′ splice donor site and the first 3′ splice acceptor site, resulting in degradation of mRNA encoding for the expressible polypeptide due to the polyA cleavage signal, and
- wherein presence of the ligand allows splicing between the 5′ splice donor site and the second 3′ splice acceptor site, resulting in expression of the expressible polypeptide due to splicing out of the polyA cleavage signal.
A nucleic acid molecule encoding a poly A aptamer polynucleotide comprising in a 5′ to 3′ direction:
- a) a 5′ splice donor site;
- b) an engineered intron with a
- a first 3′ splice acceptor site on the 3′ side of the engineered intron;
- and,
- a polyA switch comprising three ligand-binding aptamers each with one or more ligand binding pockets, a three way junction, and at least one polyA cleavage signal therein;
- wherein the first 3′ splice acceptor site is either located 5′ of the polyA switch or in the polyA switch;
- d) a second 3′ splice acceptor site;
- and
- e) a nucleic acid sequence encoding an expressible polypeptide;
- wherein absence of a ligand allows splicing between the 5′ splice donor site and the first 3′ splice acceptor site, resulting in degradation of mRNA encoding for the expressible polypeptide due to the polyA cleavage signal, and
- wherein presence of the ligand allows splicing between the 5′ splice donor site and the second 3′ splice acceptor site, resulting in expression of the expressible polypeptide due to splicing out of the polyA cleavage signal.
Dependencies are inferred from the claim text.