Patent applications filed by the University of Cincinnati

Technology overview

Dr Daniel W. Nebert at the Department of Environmental Health, University of Cincinnati conducts research on biological aspects of of metabolism-gene, receptor-gene and transporter-gene superfamilies, one direction of which includes heavy metal toxicity.  Dr Nebert filed a US patent application disclosing a method to measure water contaminants using transgenic zebrafish.  Three scientific articles were published by the research team in relation to this technology:

  1. Carvan III et al. (2000).  Transgenic zebrafish as sentinels for aquatic pollution.  Ann N Y Acad Sci. 919:133-47.
  2. Carvan III et al. (2000).  Activation of transcription factors in zebrafish cell cultures by environmental pollutants.  Arch Biochem Biophys 376: 320.
  3. Nebert et al. (2002).  Use of reporter genes and vertebrate DNA motifs in transgenic zebrafish as sentinels for assessing aquatic pollution.  Environ Health Perspect. 110(1):A15.  (Correspondence article in responce to Mattingly et al. 2001; Environ. Health Perspect. 109:845-849)

Details of the patent document

Patent or Publication no. Title, Independent Claims and Summary Assignee and licensing information
US 2004/147030

  • Earliest priority – 22 May 2000
  • Filed – 22 May 2001
  • Abandoned – 2 August 2006
  • Expected expiry – not applicable
Title – Transgenic animals for monitoring water quality

Claim 1

A method of measuring contaminants in water comprising:

a. introducing into a transgenic zebrafish organism a DNA construct having the sequence of at least one regulatory response element gene operatively linked to a DNA molecule encoding at least one reporter gene such that the at least one regulatory element of the gene controls the expression of the at least one reporter gene;
b. exposing the transgenic zebrafish to a water sample to be tested for a time sufficient to allow contaminants become bioconcentrated within the zebrafish;
c. exposing the transgenic zebrafish to conditions permitting expression of the at least one reporter gene; and
d. detecting the expression of the at least one reporter gene; and
e. correlating the detected expression to known standards and thereby determining the quantity of contaminants in the water sample.

Claim 2
A method of measuring contaminants in water comprising:a. introducing into a transgenic zebrafish organism a DNA construct having the sequence of two or more regulatory response element genes operatively linked to a DNA molecule encoding at least one reporter gene such that a regulatory elements of the gene controls expression of the reporter gene;
b. exposing the transgenic zebrafish to a water sample to be tested for a time sufficient to allow contaminants become bioconcentrated within the zebrafish
c. exposing the transgenic zebrafish to conditions permitting expression of the reporter genes; and
d. detecting the expression of the reporter genes; and
e. correlating the detected expression to known standards and thereby determining the quantity of contaminants in the water sample.

The claims are generally drawn towards:

  • a method of measuring contaminants in water comprising transgenic zebrafish with at least one (or more) regulatory response element gene(s) operatively linked to a DNA molecule encoding at least one reporter gene (claim 1, 2)

Definitions extracted from the description are:

  • Operatively linked – a juxtaposition wherein the components so described are in a relationship permitting them to function in their intended manner
  • Regulatory response element gene – those sequences normally within 100 kb of the coding region of a locus
  • Reporter gene – The present invention utilizes the firefly luciferase (luc) or green fluorescent protein (GFP)… (there is no defined statement that limits the reporter genes to luc or GFP)

Comment:

Since this is a published application and not a granted patent, currently there are no enforceable rights.

The description provides a table (Table 1) that introduces six pollution-inducible response elements and their respective consensus sequences, and an example of responses of transgenic zebrafish cell lines to substances with the following nine different pollutant-inducible promoters linked to the firefly luc gene (Figure 2):

1. Aromatic hydrocarbon response element (AHRE) to dioxin (10nM TCDD)

  • mouse Cyp1a1
  • mouse AhRDtk
  • rainbow trout CYP1A3
  • human CYP1A1

2. Electrophile response element (EPRE) to tBHQ (10µM)

  • mouse EPREmt1
  • mouse NQO1
  • human NQO1

3. Metal response element (MRE) to cadmium (30µM)

  • mouse MREd5mt1
  • rainbow trout MT-B
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US 2006/143718

  • Earliest priority – 22 May 2000
  • Filed – 23 November 2005
  • Application pending
  • Expected expiry – not applicable
Title – Transgenic animals for monitoring water quality

Claim 1

A method of measuring contaminants in water comprising:

a. introducing into a transgenic zebrafish organism a DNA construct having the sequence of at least one regulatory response element gene operatively linked to a DNA molecule encoding at least one reporter gene such that the at least one regulatory element of the gene controls the expression of the at least one reporter gene;
b. exposing the transgenic zebrafish to a water sample to be tested for a time sufficient to allow contaminants become bioconcentrated within the zebrafish;
c. exposing the transgenic zebrafish to conditions permitting expression of the at least one reporter gene; and
d. detecting the expression of the at least one reporter gene; and
e. correlating the detected expression to known standards and thereby determining the quantity of contaminants in the water sample.

Claim 2
A method of measuring contaminants in water comprising:a. introducing into a transgenic zebrafish organism a DNA construct having the sequence of two or more regulatory response element genes operatively linked to a DNA molecule encoding at least one reporter gene such that a regulatory elements of the gene controls expression of the reporter gene;
b. exposing the transgenic zebrafish to a water sample to be tested for a time sufficient to allow contaminants become bioconcentrated within the zebrafish
c. exposing the transgenic zebrafish to conditions permitting expression of the reporter genes; and
d. detecting the expression of the reporter genes; and
e. correlating the detected expression to known standards and thereby determining the quantity of contaminants in the water sample.

The claims are generally drawn towards:

  • a method of measuring contaminants in water comprising transgenic zebrafish with at least one (or more) regulatory response element gene(s) operatively linked to a DNA molecule encoding at least one reporter gene (claim 1, 2)

Definitions extracted from the description are provided in US 2004/147030 (see above).

Comments:

This application is a continuation of now abandoned US 2004/147030.

Search strategy

Search details
Date of search 13 Sept 2006
Database searched Patent Lens
Type of search Simple, stemming on
Collections searched AU-B, US-A, US-B, EP-B, WO
Search terms biomonitor and zebrafish
Results 2
Comments