Cells for drug discovery

• US 6,689,558 B2
• Filed: 02/08/2001
• Issued: 02/10/2004
• Est. Priority Date: 02/08/2000
• Status: Expired due to Term

• Current Assignee: Sangamo Biosciences, Inc.
• Original Assignee: Sangamo Biosciences, Inc.
• Inventors: Case, Casey
• Primary Examiner(s): Yucel, Remy
• Assistant Examiner(s): KATCHEVES, KONSTANTINA T
• Application Number:
  US09/779,233
• Publication Number:
  US 20020045158A1
• Time in Patent Office: 1,097 Days
• Field of Search: 435/6, 435/4, 435/7.21, 435/7.4, 435/29, 435/3
• US Class Current: 435/4
• CPC Class Codes: G01N 2500/00 Screening for compounds of potential therapeutic value
• CPC Class Codes: G01N 33/5008 for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/502 for testing non-proliferative effects
• CPC Class Codes: G01N 33/5023 on expression patterns
• CPC Class Codes: G01N 33/5041 involving analysis of members of signalling pathways
• CPC Class Codes: G01N 33/6872 Intracellular protein regulatory factors and their receptors, e.g. including ion channels

Abstract

Disclosed herein are compositions and method useful in screening a compound for its interaction and/or effect with a molecular target and/or cellular process.

Claims

1. A method of screening a compound for interaction with a molecular target comprising:

(a) contacting a first cell with the compound;

(b) determining a first value of a property of the first cell, the property being responsive to, levels of the molecular target;

(c) contacting a second cell with the compound, wherein the second cell comprises a polynucleotide encoding an exogenous zinc finger protein that directly or indirectly modulates expression of the molecular target;

(d) determining a second value of the property in the second cell, wherein a difference between the first and second values provides an indication of interaction between the compound and the molecular target.

2. The method according to claim 1, wherein the exogenous zinc finger protein directly modulates expression of the molecular target.

3. The method of claim 1, wherein the exogenous zinc finger protein further comprises a functional domain.

4. The method of claim 3, wherein the functional domain comprises an activation domain.

5. The method of claim 4, wherein the activation domain is selected from the group consisting of VP616, p65 subunit of NF-kappa B, VP64 and ligand-bound thyroid hormone receptor.

6. The method of claim 3, wherein the functional domain comprises a repression domain.

7. The method of claim 6, wherein the repression domain is selected from the group consisting of KRAB, MBD-2B, v-ErbA, MBD3, unliganded TR and members of the DNMT family.

8. The method according to claim 1, wherein the molecular target is involved in a signal transduction pathway.

9. The method according to claim 8, wherein the compound is screened for its capacity to block transduction of a signal through the molecular target.

10. The method according to claim 1, wherein the second cell comprises a second exogenous zinc finger protein.

11. The method according to claim 1, wherein the first and second cells are substantially identical except for the polynucleotide encoding the zinc finger protein.

12. The method of claim 1, wherein the polynucleotide is stably transfected into the second cell.

13. The method of claim 1, wherein the polynucleotide encoding the zinc finger protein is operably linked to an inducible promoter.

14. The method of claim 13, wherein expression of the zinc finger protein is inducted in the second cell.

15. The method of claim 13, wherein the first and second cells comprise the polynucleotide encoding the inducible zinc finger protein.

16. The method of claim 15, wherein expression of the zinc finger protein is not induced in the first cell.

17. The method according to claim 1, wherein the molecular target is a protein.

18. The method according to claim 1, wherein the molecular target is a cell surface receptor.

19. The method according to claim 1, wherein the expression level of the molecular target in the second cell is less than 25% of the expression level in the first cell.

20. The method according to claim 1, wherein the expression level of the molecular target in the second cell is more than 125% of the expression level in the first cell.

21. The method according to claim 1, wherein the expression level of the molecular target in the second cell is less than 5% of the expression level in the first cell.

22. The method according to claim 1, wherein the property is selected from the group consisting of growth, neovascularization, reporter expression and selectable marker expression.