Aptamer-mediated regulation of gene expression

Abstract

This invention provides methods of regulating gene expression. An aptamer is positioned in a nucleic acid molecule along with a sequence encoding a transcriptional regulatory polypeptide. The aptamer disrupts translation of the transcriptional regulatory polypeptide when contacted with an aptamer-binding ligand. Gene expression levels can be either increased or decreased by the disclosed methods, depending on whether the transcriptional regulatory polypeptide is a repressor or activator, and the degree of the effect is dependent upon the dose of the ligand. Nucleic acid molecules, expression cassettes, expression vectors and cells useful in the gene regulation methods are also provided.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application claims the benefit of U.S. Provisional Application No. 60 / 242,106, filed Oct. 20, 2000, which application is incorporated herein by reference for all purposes.FIELD OF THE INVENTION [0002] This invention pertains to the field of gene regulation. In particular, the invention pertains to the use of aptamers to control translation of transcriptional regulatory polypeptides. BACKGROUND OF THE INVENTION [0003] The search for therapeutics is largely focused on the regulation of gene expression and the inactivation of gene products. The development of in vitro selection techniques for identifying aptamer sequences that specifically bind a desired target molecule has provided new opportunities for the manipulation of biological interactions. Aptamers are nucleic acid molecules that are capable of binding to a particular molecule of interest with high affinity and specificity (Tuerk and Gold, Science 249:505 (1990); Ellington and...

AI Technical Summary

Benefits of technology

The present invention provides methods for regulating gene expression that are dose-responsive and can upregulate or downregulate transgenes and endogenous genes. One method involves using an aptamer-binding ligand to inhibit translation of a transcriptional regulatory polypeptide, resulting in a change in gene expression. Another method involves using a nucleic acid construct with an aptamer and a polynucleotide that encodes a transcriptional regulatory polypeptide, where the binding of a ligand to the aptamer inhibits translation of the transcriptional regulatory polypeptide, resulting in a change in gene expression and ameliorating undesirable cell proliferation. The invention also provides expression cassettes, vectors, and cells comprising the nucleic acids molecules of the invention.

Problems solved by technology

The binding of the ligand to the aptamer inhibits translation of the transcriptional regulatory polypeptide, thus causing a change in the expression level of the gene, which, in turn, ameliorates the undesirable cell proliferation.

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