Assays for screening for inhibitors of HIV
AssigneeRibotargets, Ltd.
InventorsJonathan Karn, Rodney Warren Zemmel, Peter Jonathan Gasking Butler, Roger K. Craig, Alistair Simpson Irvine
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Abstract
The invention relates to an isolated nucleic acid comprising two operatively linked binding sites for HIV Rev protein, the sites comprising a nucleation motif and an oligomerization motif, wherein the nucleic acid binds Rev protein monomers with a higher degree of co-operativity than wild-type RRE.
Claims19
A screening assay for inhibition of Rev binding to a nucleic acid substrate, comprising contacting an isolated nucleic acid comprising two operatively linked binding sites for HIV Rev protein, said sites comprising a nucleation motif and an oligomerization motif, wherein said nucleic acid binds Rev protein monomers with a higher degree of co-operativity than the wild-type RRE, with Rev protein under conditions which permit formation of a complex between said nucleic acid and Rev protein, and detecting a decrease in the amount of said complex formed in the presence of a candidate inhibitor relative to the absence of said candidate inhibitor said decrease being indicative of inhibition.
An assay for identifying a candidate inhibitor of HIV, comprising
- contacting an isolated nucleic acid comprising two operatively linked binding sites for HIV Rev protein, said sites comprising a nucleation motif and an oligomerization motif, wherein said nucleic acid binds Rev protein monomers with a higher degree of co-operativity than the wild-type RRE with Rev protein in the presence of a candidate inhibitor under conditions such that a complex is permitted to form between said nucleic acid and Rev protein, and
- determining the amount of complex formed in the presence of said candidate inhibitor, wherein a determination of a decrease in the amount of complex formed in the presence of said candidate inhibitor relative to its absence is indicative of inhibition.
An assay for identifying a candidate inhibitor of HIV, comprising
- providing an isolated nucleic acid comprising two operatively linked biding sites for HIV Rev protein, said sites comprising a nucleation motif and an oligomerization motif wherein said nucleic acid binds Rev protein monomers with a high degree of co-operativity than the wild-type RRE, Rev protein, and a candidate inhibitor
- detecting formation of a complex between said nucleic acid and Rev protein in the presence and absence of said candidate inhibitor, and
- comparing said detected formation of complex in the presence and absence of said candidate inhibitor, wherein a difference in said detected formation of complex in the presence and absence of said candidate inhibitor is indicative of inhibition.
Dependencies are inferred from the claim text.