Cell free screening assay and methods of use
AssigneeOregon Health & Science University
InventorsMaureen Hoatlin, Stacie Stone, Alexandra Sobeck, Vincenzo Costanzo, Jean Gautier, Igor Landais
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Abstract
Methods are disclosed for identifying agents for the treatment of cancer and/or Fanconi anemia using Xenopus egg cell free extracts from which the endogenous DNA has been removed. The assays and method described herein are easily adaptable to high throughput techniques for example to screen large numbers of agents as possible agents for the treatment of cancer and/or Fanconi anemia. Kits for carrying out the disclosed assays and methods also are disclosed. Agents identified by these methods are also disclosed as are methods for treating a subject with cancer and/or Fanconi anemia with these agents.
Claims26
A method for identifying an agent for the treatment of cancer or Fanconi anemia comprising:
- contacting a Xenopus egg cell free extract depleted of endogenous DNA with an agent of interest;
- detecting a post-translationally modified Xenopus Fanconi anemia polypeptide in the Xenopus egg cell free extract;
- and
- comparing the amount of the post-translationally modified Xenopus Fanconi anemia polypeptide to a control, wherein a difference in the amount of the post-translationally modified Xenopus Fanconi anemia polypeptide in the cell extract contacted with the agent, as compared to the control, indicates the agent is of use for treating cancer, Fanconi anemia, or a combination thereof.
A method for identifying an agent for the treatment of cancer or Fanconi anemia comprising:
- contacting a Xenopus egg cell free extract depleted of endogenous DNA with an agent of interest;
- adding isolated exogenous double stranded DNA comprising at least one double strand break, wherein the exogenous double stranded DNA is between 10 and 500 nucleotides in length;
- detecting a post-translationally modified Xenopus Fanconi anemia polypeptide in the Xenopus egg cell free extract, wherein the post-translational modification comprises ubiquitination or phosphorylation;
- and
- comparing the amount of the post-translationally modified Xenopus Fanconi anemia polypeptide to a control, wherein a difference in the amount of the post-translationally modified Xenopus Fanconi anemia polypeptide in the cell extract contacted with the agent, as compared to the control, indicates the agent is of use for treating cancer, Fanconi anemia, or a combination thereof.
Dependencies are inferred from the claim text.