Whole-body optical imaging of gene expression and uses thereof

• US 6,649,159 B2
• Filed: 03/19/2001
• Issued: 11/18/2003
• Est. Priority Date: 03/17/2000
• Status: Expired due to Fees

• Current Assignee: Anticancer Incorporated
• Original Assignee: Anticancer Incorporated
• Inventors: Yang, Meng, Baranov, Eugene
• Primary Examiner(s): Falk, Anne-Marie
• Assistant Examiner(s): QIAN, CELINE X
• Application Number:
  US09/812,710
• Publication Number:
  US 20020013954A1
• Time in Patent Office: 974 Days
• Field of Search: 800/8, 435/320.1, 435/325, 536/24.5, 424/93.1
• US Class Current: 424/93.21
• CPC Class Codes: C12Q 1/6897 involving reporter genes operably linked to promoters

Abstract

The invention relates to the whole-body external optical imaging of gene expression. Specifically, methods for whole-body external optical imaging of gene expression and methods for evaluating a candidate protocol or drug for treating diseases or disorders using a fluorophore operatively linked to the promoter of a gene and external optical imaging are provided herein. Methods to screen for substances or genes that regulate target promoters are also provided.

Claims

1. A method to monitor the ability of a promoter to promote expression in an animal of an endogenous gene that is controlled by said promoter, which method comprises:

a) delivering, to an animal, cells containing a nucleic acid encoding a fluorophore operatively linked to the promoter of said endogenous gene whose ability to promote expression is to be analyzed; and

b) observing the presence, absence or intensity of the fluorescence generated by said fluorophore at various locations in said animal by whole-body external fluorescent optical imaging,

whereby the ability of said promoter to promote expression is monitored, and

wherein said fluorophore is a protein that is autofluorescent such that no substrates or cofactors are needed for it to fluoresce.

2. The method of claim 1, wherein the cells are delivered to the animal via a surgical procedure.

3. The method of claim 2, wherein the cells are delivered to the animal via direct implantation by surgical orthotopic implantation (SOI) at a desired site.

4. The method of claim 1, wherein the animal is a human and the fluorophore is a humanized fluorophore.

5. The method of claim 1, wherein the fluorophore is selected from the group consisting of a green fluorescent protein (GFP), a blue fluorescent protein (BFP) and a red fluorescent protein (RFP).

6. The method of claim 5, wherein the animal is a human and the GFP is the humanized hGFP-S65T.

7. The method of claim 1, wherein the animal is a mammal.

8. The method of claim 7, wherein the mammal is selected from the group consisting of a mouse, a rat, a rabbit, a cat, a dog, a pig, a cow, an ox, a sheep, a goat, a horse, a monkey and a non-human primate.

9. The method of claim 1, wherein the endogenous gene is normally expressed in a tissue or organ specific manner.

10. The method of claim 9, wherein the tissue is selected from the group consisting of connective, epithelium, muscle and nerve tissues.

11. The method of claim 9, wherein the organ is selected from the group consisting of brain, lung, liver, spleen, bone marrow, thymus, heart, lymph, blood, bone, cartilage, pancreas, kidney, gall bladder, stomach, intestine, testis, ovary, uterus, rectum, nervous system, gland, and internal blood vessels.

12. The method of claim 1, wherein the endogenous gene is an endogenous tumor or cancer associated gene.

13. The method of claim 12, wherein the tumor or cancer associated gene is an oncogene or a tumor suppressor gene.