Methods and compositions for screening for angiogenesis modulating compounds

A method for monitoring expression of an angiogenesis-related gene in a living transgenic rodent, said method comprising:

• monitoring the expression of a reporter sequence in a living transgenic rodent, said rodent comprising an expression cassette, said expression cassette comprising
• (a) a transcription control element derived from an angiogenesis-related gene, wherein said transcription control element is selected from the group consisting of
• (I) a VEGFR-2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,564 of SEQ ID NO:32, (ii) X equals Y, and (iii) X is greater than or equal to 55;
• (II) a VEGF-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,762 of SEQ ID NO:44, (ii) X equals Y, and (iii) X is greater than or equal to 50;
• and
• (III) a TIE2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (A) (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 6,091 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 55, or (B) (i) the X contiguous nucleotides have at least about 90% identity to Y contiguous nucleotides derived from nucleotides 6,091 through 6,560 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 250;
• and
• (b) a reporter sequence operably linked to said transcription control element, wherein said reporter sequence encodes a light generating protein, and wherein expression of said reporter sequence indicates expression of said angiogenesis-related gene.

A method for monitoring expression of an angiogenesis-related gene in a cell, said method comprising

• monitoring the expression of a reporter sequence in a cell, said cell comprising an expression cassette, said expression cassette comprising
• (a) a transcription control element derived from an angiogenesis-related gene, wherein said transcription control element is selected from the group consisting of:
• (I) a VEGFR-2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,564 of SEQ ID NO:32, (ii) X equals Y, and (iii) X is greater than or equal to 55;
• (II) a VEGF-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,762 of SEQ ID NO:44, (ii) X equals Y, and (iii) X is greater than or equal to 50;
• and
• (III) a TIE2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (A) (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 6,091 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 55, or (B) (i) the X contiguous nucleotides have at least about 90% identity to Y contiguous nucleotides derived from nucleotides 6,091 through 6,560 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 250;
• and
• (b) a reporter sequence operably linked to said transcription control element, wherein said reporter sequence encodes a light-generating protein, and wherein expression of said reporter sequence indicates expression of said angiogenesis-related gene.

A method for identifying a compound which affects angiogenesis, said method comprising

• (a) administering a compound to a living transgenic rodent, said rodent comprising an expression cassette, said expression cassette comprising
• (i) a transcription control element derived from an angiogenesis-related gene, wherein said transcription control element is selected from the group consisting of:
• (I) a VEGFR-2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,564 of SEQ ID NO:32, (ii) X equals Y, and (iii) X is greater than or equal to 55;
• (II) a VEGF-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,762 of SEQ ID NO:44, (ii) X equals Y, and (iii) X is greater than or equal to 50;
• and
• (III) a TIE2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (A) (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 6,091 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 55, or (B) (i) the X contiguous nucleotides have at least about 90% identity to Y contiguous nucleotides derived from nucleotides 6,091 through 6,560 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 250;
• and
• (ii) a reporter sequence operably linked to said transcription control element, wherein said reporter sequence encodes a light-generating protein, and wherein expression of said reporter sequence indicates expression of said angiogenesis-related gene;
• and
• (b) monitoring the expression of said reporter sequence wherein an effect on the expression of said reporter sequence mediated by said compound indicates that the compound affects angiogenesis.

A method for identifying a compound which affects angiogenesis, said method comprising

• (a) administering a compound to a cell, said cell comprising an expression cassette, said expression cassette comprising
• (i) a transcription control element derived from a gene involved in angiogenesis, wherein said transcription control element is selected from the group consisting of
• (I) a VEGFR-2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,564 of SEQ ID NO:32, (ii) X equals Y, and (iii) X is greater than or equal to 55;
• (II) a VEGF-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 3,762 of SEQ ID NO:44, (ii) X equals Y, and (iii) X is greater than or equal to 50;
• and
• (III) a TIE2-locus-derived sequence comprising a polynucleotide having X contiguous nucleotides, wherein (A) (i) the X contiguous nucleotides have at least about 95% identity to Y contiguous nucleotides derived from nucleotides 1 through 6,091 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 55, or (B) (i) the X contiguous nucleotides have at least about 90% identity to Y contiguous nucleotides derived from nucleotides 6,091 through 6,560 of SEQ ID NO:40, (ii) X equals Y, and (iii) X is greater than or equal to 250;
• and
• (ii) a reporter sequence operably linked to said transcription control element, wherein said reporter sequence encodes a light-generating protein;
• and
• (b) monitoring the expression of said reporter sequence wherein an effect on the expression of said reporter sequence mediated by said compound indicates that the compound affects angiogenesis.