SEQUENCE-SPECIFIC INHIBITION OF SMALL RNA FUNCTION

摘要

The present invention relates to the discovery of a method for inhibiting RNA silencing in a target sequence-specific manner. RNA silencing requires a set of conserved cellular factors to suppress expression of gene-encoded polypeptide. The invention provides compositions for sequence-specific inactivation of the RISC component of the RNA silencing pathway, and methods of use thereof. The RISC inactivators of the present invention enable a variety of methods for identifying and characterizing miRNAs and siRNAs, RISC-associated factors, and agents capable of modulating RNA silencing. Therapeutic methods and compositions incorporating RISC inactivators and therapeutic agents identified through use of RISC inactivators are also featured.

主权项

1. A method for inhibiting RNA silencing of a gene, comprising, contacting a cell or organism containing an siRNA that directs RNA silencing of the gene, with a single-stranded, nuclease resistant RNA oligonucleotide RISC inactivator, wherein the RISC inactivator comprises a nucleotide sequence sufficiently complementary to a guide strand sequence of the siRNA to inhibit RNA silencing of the gene by the siRNA, and wherein the RISC inactivator is a stoichiometric, irreversible inhibitor of RISC function, such that RNA silencing of the gene is inhibited.