Non-invasive evaluation of physiological response in a transgenic mouse

• US 7,449,615 B2
• Filed: 12/16/1999
• Issued: 11/11/2008
• Est. Priority Date: 12/17/1998
• Status: Expired due to Fees

• Current Assignee: Xenogen Corporation (Revvity, Inc.)
• Original Assignee: Xenogen Corporation (Revvity, Inc.)
• Inventors: Zhang, Ning, Contag, Pamela R.
• Primary Examiner(s): FALK, ANNE MARIE
• Application Number:
  US09/464,795
• Publication Number:
  US 20020138855A1
• Time in Patent Office: 3,253 Days
• Field of Search: 800/3, 800/8, 800/13, 800/18, 800/21, 800/14
• US Class Current: 800/3
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2217/20 Animal model comprising regulated expression system
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 2267/0393 Animal model comprising a reporter system for screening tests
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: A01K 67/0276 Knock-out vertebrates
• CPC Class Codes: A01K 67/0278 Knock-in vertebrates, e.g. humanised vertebrates
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 15/8509 for producing genetically modified animals, e.g. transgenic

Abstract

The present invention relates to panels of reporter expression cassettes and the generation of transgenic non-human animals, wherein said reporter expression cassettes have selected control elements operable linked to reporter genes. The invention includes methods of use thereof for the identification and characterization of the effects of compounds administered to the live transgenic non-human animals.

Claims

1. A transgenic mouse comprising a panel of expression cassettes, said transgenic mouse produced by a method comprising the steps of

introducing a first expression cassette comprising a first promoter derived from a first stress-inducible gene into a mouse at an embryonic stage, said promoter operably linked to sequences encoding a first light generating polypeptide, and

introducing a second expression cassette comprising a second promoter derived from a second stress-inducible gene into said mouse at an embryonic stage, said promoter operably linked to sequences encoding a second light generating polypeptide and said second promoter derived from a different stress-inducible gene than said first promoter.

2. A method of determining the effect of an analyte on gene expression mediated by promoters derived from stress-inducible genes, wherein said expression is in a living transgenic mouse, said method comprising

administering the analyte to a living transgenic mouse of claim 1, wherein administering of said analyte is carried out under conditions that permit light generation mediated by said light generating polypeptide in the transgenic mouse,

determining the effect of the analyte on expression of the light generating polypeptide in a living transgenic mouse wherein said expression is mediated by at least one of the promoters.

3. The method of claim 2, wherein said conditions that permit light generation mediated by the light generating polypeptide includes administering, to the transgenic mouse, at least one substrate for the light generating polypeptide.

4. A noninvasive method for detecting a level of expression in response to an analyte, wherein said expression is (i) mediated by promoters derived from stress-inducible genes, and (ii) in a living transgenic mouse, said method comprising

(a) administering the analyte to a living transgenic mouse of claim 1, wherein administering of said analyte is carried out under conditions that permit light generation mediated by said light generating polypeptide,

(b) placing the transgenic mouse within a detection field of a photo detector device,

(c) maintaining the transgenic mouse in the detection field of the device, and

(d) during said maintaining, measuring photon emission from the transgenic mouse with the photo detector device to detect the level of expression of the light generating polypeptide in the living transgenic mouse wherein said expression is mediated by at least one of the promoters.

5. The method of claim 4, further comprising, (e) repeating steps (b) through (d) at selected intervals, wherein said repeating is effective to detect changes in the level of the light emission in the transgenic mouse over time.

6. A method of providing a transgenic mouse suitable for screening a selected analyte, comprising

generating a transgenic mouse of claim 1, and

providing said transgenic mouse or progeny thereof for use in screening a selected analyte.

7. The transgenic mouse of claim 1, wherein the method further comprises

introducing a third expression cassette comprising a promoter derived from a third stress-inducible gene into a mouse at an embryonic stage, said third promoter operably linked to sequences encoding a third light generating polypeptide and said third promoter derived from a different stress-inducible gene than said first and second promoters.

8. The transgenic mouse of claim 7, wherein (i) said first, second, and third promoters are each derived from a different gene, and (ii) said first, second, and third light generating polypeptides produce the same color of light.

9. The transgenic mouse of claim 7, wherein (i) said first, second, and third promoters are each derived from a different gene, and (ii) at least two of said first, second, and third light generating polypeptides produce different colors of light.

10. The transgenic mouse of claim 7, said panel further comprising additional expression cassettes, wherein each expression cassette comprises a promoter derived from a different stress-inducible gene, said promoter operably linked to sequences encoding a light generating polypeptide.