Wisconsin Alumni Research Foundation

The Royal Institution for the Advancement of Learning / Mcgill University

Other

The State University of New York

The University of Arizona

Marvin P Wickens

Stanley Fields

Franco Cerrina

Masad Jose Damha

Jeremy Lackey

Roy Parker

Mark Somoza

Debbie Mitra

Melanie A Preston

Michael D Sheets

Christopher P Lapointe

Masad Damha

Methods, kits, and compositions of matter relating to poly(UG) polymerases are disclosed. In one embodiment, a method includes: contacting an RNA substrate with a poly(UG) polymerase; and allowing the poly(UG) polymerase to add a poly(UG) sequence to the end of the RNA substrate by retaining contact between the RNA substrate and the poly(UG) polymerase for a period of time from about 1 second to about 28 days. The poly(UG) polymerase can beRDE-3.

Poly(UG) polymerase, constructs, and methods of making and using the same

The present invention is directed to RNA monomers comprising O-acetal levulinyl protecting groups at the 2' and/or the 5&#x2032;-hydroxy functionalities of the ribose moiety. Said monomers may be incorporated into oligoribonucleotides or RNA polynucleotides. Furthermore, the invention is directed to methods for the synthesis of said RNA monomers, oligoribonucleotides and RNA polynucleotides, as well as methods for their deprotection and methods for the use of said compounds and compositions comprising said compounds. In particular, such compounds and compositions comprising them are used in methods for light-directed synthesis of RNA microarrays.

RNA monomers containing 0-acetal levulinyl ester groups and their use in RNA microarrays

Methods, kits, and compositions of matter suitable for use in RNA Tagging are disclosed. In one embodiment, a method includes: expressing a fusion protein within the cellular environment, the fusion protein including at least part of the protein of interest and a tagging domain, the tagging domain introducing a selective tag to an RNA to which the fusion protein selectively binds, the selective tag including a selective tag sequence or a selective covalent modification; allowing the tagging domain to tag the RNA to which the protein of interest selectively binds by waiting for about 1 minute to about 28 days; and identifying the tagged RNA.

RNA tagging

RNA monomers containing O-acetal levulinyl ester groups and their use in RNA microarrays

A method for determining the function of a test protein is disclosed. In a preferred embodiment, this method comprises the steps of fusing a test protein to an RNA-binding protein and exposing the fusion protein to recombinant reporter mRNA molecule that comprises a binding site for the RNA-binding protein in the 3&prime; untranslated region. One then observes the properties of reporter mRNA and correlates the reporter properties with test protein function.

Tethered function assay for protein function

A method for determining the function of a test protein is disclosed. In a preferred embodiment, this method comprises the steps of fusing a test protein to an RNA-binding protein and exposing the fusion protein to recombinant reporter mRNA molecule that comprises a binding site for the RNA-binding protein in the 3' untranslated region. One then observes the properties of reporter mRNA and correlates the reporter properties with test protein function.

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.

System to detect protein-RNA interactions

Translation characteristics of an mRNA molecule in a cell can be altered by providing a prosthetic RNA molecule that includes a regulatory sequence element and a sequence element complementary to a portion of the mRNA molecule. Alteration can affect the translation rate, the mRNA stability, or the localization of the mRNA molecule.

Growing community of inventors

Madison, WI, United States of America

Marvin P Wickens