Precursor miRNA loop-modulated target regulation

Patent metadata

Field	Value
Publication number	US-9388466-B2
Application number	US-201414457656-A
Country	US
Kind code	B2
Filing date	Aug 12, 2014
Priority date	Jun 22, 2007
Publication date	Jul 12, 2016
Grant date	Jul 12, 2016

Abstract

Modulation of mRNA activity is achieved with precursor miRNAs (ta-RNAs). ta-RNAs, primarily pre-miRNAs and pri-miRNAs, including truncated and mutated ta-RNAs, are employed for modulation of mRNA expression where it is found that pri- and pre-miRNA have activity independently of the presence of functional mature miRNAs. Modification of at least one of the stem and loop of the ta-RNAs to enhance binding of the ta-RNA to the target mRNA is employed. The modification may be enhanced complementarity between the ta-RNA and the target mRNA and/or improved thermodynamic efficiency in binding of the ta-RNA to the target.

First claim

What is claimed is:

1. A method for identifying the role of a loop sequence of a precursor miRNA among a plurality of isoforms differing in their loop sequences, said method comprising:
substituting one loop from one isoform with an equivalent loop from a different isoform to produce a mutated precursor miRNA;
introducing said one isoform in a first cell and said mutated precursor miRNA into a second cell;
determining the effect of the one isoform and said mutated precursor miRNA on the phenotype of each of said first and second cells;
wherein said phenotype is different, indicating that said loop sequence affects the role of said precursor miRNA; and
identifying said loop sequence as affecting the role of said precursor miRNA.

2. The method according to claim 1 , wherein said first and second cells are T-cells and said phenotype is the differentiation of said T-cells.

Assignees

• Univ Leland Stanford Junior
• Leland Stanford Jr Univ Board Of Regents

Inventors

• Chen Chang-Zheng
• Trujillo Robin
• Yue Sibiao

Classifications

• C12N2501/515

  CD3, T-cell receptor complex · CPC title

• C12N15/111 Primary

  General methods applicable to biologically active non-coding nucleic acids · CPC title

• C12N2501/42

  Notch; Delta; Jagged; Serrate · CPC title

• C12Q1/6811

  Selection methods for production or design of target specific oligonucleotides or binding molecules · CPC title

• C12N2506/11

  from blood or immune system cells · CPC title