System to detect protein-RNA interactions

US5677131A · kind A · utility

Assignee

• Wisconsin Alumni Research Foundation · US

Inventors

• Marvin P. Wickens · Madison, US
• Stanley Fields · East Setauket, US

Key dates

• Filing date: Jul 9, 1996
• Grant date: Oct 14, 1997
• Expiry date: Jul 9, 2016

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N2740/16322
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A method for detecting an interaction between an RNA-binding protein and a test RNA molecule is disclosed. This method comprises providing a host cell containing a detectable gene. The detectable gene expresses a detectable protein when the detectable gene is activated by an amino acid sequence including a transcriptional activation domain when the transcriptional activation domain is in sufficient proximity to the detectable gene. First, second and third chimeric genes are also provided. The first chimeric gene comprises a DNA-binding domain that recognizes a binding site on the detectable gene in the host cell and a first RNA-binding domain. The second chimeric gene comprises a transcriptional activation domain and a second RNA-binding domain. The third chimeric gene comprises a first RNA sequence capable of binding to either the first or second RNA-binding and a second RNA sequence to be tested for interaction with the RNA-binding protein not bound to the first RNA sequence. Interaction between both the first RNA-binding domain and the hybrid RNA and the second RNA-binding domain and the hybrid RNA in the host cell causes expression of the detectable gene.