System to detect protein-RNA interactions

Details System to detect protein-RNA interactions System to detect protein-RNA interactions

1996-11-12

1998-05-12

Ketter, James

Organic compounds -- part of the class 532-570 series

Organic compounds

Carbohydrates or derivatives

435 6, C07H 2102, C12Q 168

Patent

active

057506671

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.

REFERENCES:
patent: 5283173 (1994-02-01), Fields et al.
Chien, C.-T., et al., "The two-hybrid system: A method to identify and clone genes for proteins that interact with a protein of interest," Proc. Natl. Acad. Sci. USA 88:9578-9582, 1991.
Fields, S., et al., "A novel genetic system to detect protein-protein interactions," Nature 340:245-246, 1989.
Frankel, A.D., et al., "RNA-Protein Interactions," Cell 67:1041-1046, 1991.
"Introducing the Matchmaker Two-Hybrid System," Clontech Advertisement, date unkown.
Spencer et al. Conrolling signal transduction with synthetic ligands Science vol. 262 1019-1024, 1993.
Silar et al. New shuttle vectors for direct cloning in Saccharomyces cerevisiae Gene vol. 104 99-102, 1991.
Stripecke et al. Proteins binding to 5' untranslated region sites: a general mechanism for translational regulation of mRNAs in human and yeast cells Mol. Cell. Biol. vol. 14 5898-5909, 1994.
Mattaj RNA recognition: A family matter? Cell vol. 73 837-840, 1993.
Fields et al. The two-hybrid system: an assay for protein-protein interactions Trends in Genetics vol. 10 286-292, 1994.
Good et al. Yeast expression vectors using RNA polymerase III promoters Gene vol. 151 209-214, 1994.

Affiliated with

Also associated with

Rating

System to detect protein-RNA interactions does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with System to detect protein-RNA interactions, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and System to detect protein-RNA interactions will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-980120