Isolation and identification of transcription control elements associated with mouse eosinophil peroxidase expression
AssigneeXenogen Corporation
InventorsHongbing Zhang, Pamela Contag, Anthony Purchio, Amy Holt
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Abstract
The present invention relates to novel transcription control elements derived from a mouse eosinophil peroxidase gene. Such transcription control elements may comprise isolated polynucleotides, expression cassettes, vectors, recombinant cells, and transgenic animals, as described herein.
Claims6
A method for identifying an analyte that modulates expression of a sequence encoding a light-generating protein, wherein expression of said light-generating protein is mediated by transcription control elements derived from a mouse eosinophil peroxidase gene, in a living transgenic mouse, said method comprising
- providing a living transgenic mouse whose genome comprises an expression cassette comprising a polynucleotide having nucleotides 1 to 9,825 of SEQ ID NO:1 or a variant thereof that have at least 95% or greater identity to nucleotides 1–9,825 of SEQ ID NO:1 over the entire length of nucleotides 1–9,825, operably linked to a sequence encoding a light-generating protein, wherein the polynucleotide causes increased expression of the light-generating protein in eosinophils of the living transgenic mouse as compared to non-eosinophil blood cell-types;
- administering to the living transgenic living mouse said analyte;
- and
- monitoring expression of said light-generating protein wherein an effect on the level of expression of said light-generating protein indicates that the analyte affects expression mediated by transcription control elements derived from the mouse eosinophil peroxidase gene.
A method for monitoring eosinophil cell location in a living transgenic mouse, said method comprising
- providing a living transgenic mouse whose genome comprises an expression cassette comprising a polynucleotide having nucleotides 1 to 9,825 of SEQ ID NO:1 or a variant thereof that have at least 95% or greater identity to nucleotides 1–9,825 of SEQ ID NO:1 over the entire length of nucleotides 1–9,825, operably linked to a sequence encoding a light-generating protein, wherein the polynucleotide causes increased expression of the light-generating protein in eosinophils of the living transgenic mouse as compared to non-eosinophil blood cell-types;
- inducing eosinophil production in the living transgenic mouse, and
- monitoring eosinophil cell location in the living transgenic mouse by monitoring locations of expression of the light-generating protein in regions of the body of the living transgenic mouse.
A method for evaluating the effect of an analyte on eosinophil migration in a living transgenic mouse, said method comprising
- providing a living transgenic mouse whose genome comprises an expression cassette comprising a polynucleotide having nucleotides 1 to 9,825 of SEQ ID NO:1 or a variant thereof that have at least 95% or greater identity to nucleotides 1–9,825 of SEQ ID NO:1 over the entire length of nucleotides 1–9,825, operably linked to a sequence encoding a light-generating protein, wherein the polynucleotide causes increased expression of the light-generating protein in eosinophils of the living transgenic mouse as compared to non-eosinophil blood cell-types;
- inducing eosinophil migration to a selected site in first and second living transgenic mice,
- administering to the first living transgenic mouse an analyte,
- monitoring eosinophil migration to the selected site in the first and second living transgenic mice by monitoring expression of the light-generating protein in the living transgenic mice, and
- evaluating any effect of the analyte on eosinophil migration by comparing eosinophil migration in said first and second living transgenic mice.
A method for inducing eosinophil, cell-type specific expression of a coding sequence of interest in a living transgenic mouse, comprising
- inducing eosinophil production in a living transgenic mouse whose genome comprises an expression cassette comprising a polynucleotide having nucleotides 1 to 9,825 of SEQ ID NO:1 or a variant thereof that have at least 95% or greater identity to nucleotides 1–9,825 of SEQ ID NO:1 over the entire length of nucleotides 1–9,825, operably linked to a sequence encoding a light-generating protein, wherein the polynucleotide causes increased expression of the light-generating protein in eosinophils of the living transgenic mouse as compared to non-eosinophil blood cell-types;
- wherein said induction of eosinophil production results in eosinophil, cell-type specific expression of the light-generating protein in the living transgenic mouse.
Dependencies are inferred from the claim text.