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Abstract
Grant Number: 1R03NS056978-01 Project Title: Screen for molecules inhibiting histone pre-mRNA process
PI Information: Name Title MARZLUFF, WILLIAM F. marzluff@med.unc.edu PROFESSOR Abstract: DESCRIPTION (provided by applicant): We will develop a cell-based assay that allows us to detect changes in the ability to process histone pre- mRNA. The reporter encodes a portion of a histone mRNA including the 3' end formation signal fused to a downstream GFP open reading frame followed by a polyadenylation signal. Failure to process at the histone 3' end will result in read-through to the polyadenylation site resulting in production of GFP. We have successfully carried out a genome-wide RNA interference screen using this approach. We will adapt the screen to screening for chemical compounds. Using an antisense oligonucleotide which blocks U7 snRNP, we have shown that we can readily detect the inhibition of histone pre-mRNA processing by fluorescence of the reporter. We are adapting the screen to an automated microscope which can rapidly read the 384 well plates.
Public Health Relevance:
This Public Health Relevance is not available.Thesaurus Terms:
RNA biosynthesis, RNA interference, drug screening /evaluation, histone, posttranscriptional RNA processing, precursor mRNA
antisense nucleic acid, green fluorescent protein, high throughput technology, oligonucleotide, reporter gene
NIH Roadmap Initiative tag, tissue /cell culture
Institution: UNIVERSITY OF NORTH CAROLINA CHAPEL HILL Office of Sponsored Research CHAPEL HILL, NC 27599 Fiscal Year: 2006 Department: BIOCHEMISTRY AND BIOPHYSICS Project Start: 01-JUL-2006 Project End: 30-JUN-2007 ICD: NATIONAL INSTITUTE OF NEUROLOGICAL DISORDERS AND STROKE IRG: ZNS1