Screen for CDC7 inhibitors
AssigneeMillennium Pharmaceuticals, Inc.
InventorsBenjamin Stone Amidon, Christine Ellen Bulawa
Life of this patent
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Abstract
Disclosed is a yeast cell whose genetic complement includes an inactive allele of the yeast CDC7 gene, a first nucleic acid that encodes a mammalian Cdc7 protein, and a second nucleic acid that encodes a mammalian Dbf4 protein. The yeast cell is dependent on the mammalian Cdc7 and Dbf4 proteins for viability. The yeast cell can be used to identify potential anti-proliferative agents by virtue of their inhibition of the mammalian Cdc7 and Dbf4 proteins. In some embodiments, a control yeast cell which does not depend on Cdc7 for viability is used in a secondary screen.
Claims28
A yeast cell, the genetic complement of which comprises:
- a) an inactive allele of the yeast CDC7 gene;
- b) a first nucleic acid molecule that encodes a mammalian Cdc7 polypeptide;
- and
- c) a second nucleic acid molecule that encodes a mammalian Dbf4 polypeptide.
A method for determining whether a test compound is an inhibitor of mammalian Cdc7.Dbf4 activity, the method comprising:
- (i) contacting a test compound to a first yeast cell, the genetic complement of which comprises:
- (a) an inactive allele of the yeast CDC7 gene;
- (b) a first nucleic acid molecule that encodes a mammalian Cdc7 polypeptide;
- and
- (c) a second nucleic acid molecule that encodes a mammalian Dbf4 polypeptide;
- (ii) contacting the test compound to a second yeast cell, the viability of which is independent of a mammalian Cdc7 or Dbf4;
- and
- (iii) assessing growth or cell cycle progression of the first and second yeast cells, wherein a decrease in growth or cell cycle progression in the first yeast cell relative to the second yeast strain indicates that the test compound is an inhibitor of mammalian Cdc7.Dbf4 activity.
A kit comprising:
- (1) a first yeast cell, the genetic complement of which comprises a) an inactive allele of the yeast CDC7 gene, b) a first nucleic acid that encodes a mammalian Cdc7 polypeptide, and c) a second nucleic acid that encodes a mammalian Dbf4 polypeptide;
- and
- (2) a second yeast cell, the genetic complement of which comprises a) an inactive allele of the yeast CDC7 gene, and b) a mutant allele that bypasses the requirement for Cdc7.Dbf4 activity.
Dependencies are inferred from the claim text.