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US 7,598,079 B2Granted
Assay for identifying compounds which affect stability of mRNA
AssigneeNovation Pharmaceuticals Inc.
InventorsTania Kastelic, Dominique Cheneval
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Abstract
The present invention relates to an assay for the identification of biologically active compounds, in particular to a reporter gene assay for the identification of compounds, which have an effect on mRNA stability. More particularly, the present invention relates to a reporter gene expression system and cell lines comprising said expression system. The invention further relates to compounds which destabilize mRNA.
Claims21
A stably transfected cell line comprising:
- i) a DNA expression vector comprising a first DNA sequence encoding a first protein having a detectable signal, one or more 3′ UTR sequence and one or more expression control sequence operatively associated with said first DNA sequence, and a heterologous instability sequence DNA that is heterologous to the 3′UTR sequence and inserted into the 3′UTR sequence, said instability sequence DNA comprising a second DNA sequence corresponding to one or more mRNA instability sequence which is from one or more naturally occurring gene sequences;
- and ii) a control DNA expression vector comprising a control DNA sequence encoding a second protein having a detectable signal, and one or more 3′ UTR sequence and one or more expression control sequence operatively associated with said control DNA sequence, wherein said cell line is used for screening compounds which affect mRNA stability, wherein said heterologous instability sequence DNA is from about 200 to about 1500 nucleotides in length and comprises DNA corresponding to sequences that flank said mRNA instability sequence in the naturally occurring gene.
A set of stably transfected cell lines comprising:
- (i) a stably transfected cell line comprising a DNA expression vector comprising a first DNA sequence encoding a first protein having a detectable signal, one or more 3′ UTR sequence and one or more expression control sequence operatively associated with said first DNA sequence, and a heterologous instability sequence DNA that is heterologous to the 3′UTR sequence and inserted into the 3′UTR sequence, said instability sequence DNA comprising a second DNA sequence corresponding to one or more mRNA instability sequence which is from one or more naturally occurring gene sequences;
- and (ii) a stably transfected cell line comprising a control DNA expression vector comprising a control DNA sequence encoding a second protein having a detectable signal, and one or more 3′ UTR sequence and one or more expression control sequence operatively associated with said control DNA sequence, wherein said set of cell lines is used for screening compounds which affect mRNA stability, wherein said heterologous instability sequence DNA is from about 200 to about 1500 nucleotides in length and comprises DNA corresponding to sequences that flank said mRNA instability sequence in the naturally occurring gene.
Dependencies are inferred from the claim text.