Patent ID: 8283115

Claim:
A method for screening for a compound that modulates human UTRN mRNA translation that is regulated by the untranslated regions (UTRs) of the human UTRN mRNA, the method comprising: (a) contacting a compound with a first host cell engineered to express a first reporter protein translated from a first mRNA transcript comprising a first reporter gene coding sequence operably linked to a first 5′-UTR of human UTRN encoded by the nucleotide sequence of SEQ ID NO:5 or a fragment thereof and a first 3′-UTR of human UTRN encoded by the nucleotide sequence of SEQ ID NO:6 or a fragment thereof, wherein the first 5′-UTR or a fragment thereof is upstream of the nucleotide sequence of the reporter gene and the first 3′-UTR or a fragment thereof is downstream of the reporter gene coding sequence, and wherein the first reporter protein is not UTRN; and (b) contacting the compound with a second host cell engineered to express a second reporter protein translated from a second mRNA transcript comprising the first reporter gene coding sequence operably linked to a second 5′-UTR or a fragment thereof and a second 3′-UTR or a fragment thereof of a mRNA different from the first 5′-UTR of human UTRN mRNA or a fragment thereof and the first 3′-UTR of human UTRN mRNA or a fragment thereof, wherein the second 5′-UTR is upstream of the first reporter gene coding sequence and the second 3′ UTR is downstream of the first reporter gene coding sequence; and (c) detecting the amount or activity of the first and second reporter protein, wherein (i) an alteration in the amount or activity of the first reporter protein in the presence of the compound relative to the amount or activity in the absence of the compound or the presence of a negative control, and (ii) no alteration in or not a substantially altered amount or activity of the second reporter protein in the presence of the compound relative to the amount or activity of the second reporter protein in the absence of the compound or the presence of the negative control indicates that the compound modulates human UTRN mRNA translation that is regulated by the UTRs of human UTRN mRNA; and wherein the amount or activity of the first reporter protein in the presence of the compound is compared to the amount or activity of the first reporter protein in the presence of 4-(4-aminophenylthio)-6-chloropyrimidin-2-amine, N,N-dimethyl-4-(5-nitro-1H-benzo[d]imidazol-2-yl)aniline, 1-chloro-3-propyl-benzoimidazo[1,2-a]pyridine-4-carbonitrile, 3-(2,3-dihydro-1H-benzo[f]cyclopenta[c]quinolin-4-yl)phenol, N-(4-(benzo[d]thiazol-2-yl)phenylcarbamothioyl)-4-ethoxy-3-nitrobenzamide, 2-amino-4-(3-(trifluoromethyl)phenyl)-4H-benzo[h]chromene-3-carbonitrile, or N-(5-(benzo[d]thiazol-2-yl)-2-methylphenylcarbamothioyl)-4-butoxybenzamide.