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US 10,246,498 B2Granted
Genetically encoded histone reporter allele constructs
AssigneeThe Johns Hopkins University
InventorsHans Tomas Bjornsson, Harry Dietz
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Abstract
Non-FRET-based fusion protein reporter molecules are provided that can be used to monitor histone modifications in living cells. Transgenic animals, particularly non-human mammals, whose genomes comprise an expression cassette encoding a non-FRET-based fusion protein reporter, are also provided. Methods of using the fusion reporter molecules for diagnosing histone-modification-associated disorders and to identify candidate pharmaceutical agents that effect histone modification in cells and tissues are also provided.
Claims27
A fusion protein reporter comprising:
- a) a histone-modification-specific binding domain that is configured to bind to said histone polypeptide substrate when said histone polypeptide substrate is modified;
- b) a histone polypeptide substrate;
- and
- c) a flexible linker region, wherein the flexible linker region is flanked on one side by a region encoding a C-terminal portion of a circularly permutated fluorescent protein and on the other side by a region encoding the N-terminal portion of the circularly permutated fluorescent protein, and wherein said C-terminal portion of said circularly permutated fluorescent protein and said N-terminal portion of said circularly permutated fluorescent protein form a functional fluorescent protein when said histone-modification-specific binding domain binds to said histone polypeptide substrate.
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Dependencies are inferred from the claim text.