STROMAL INTERACTING MOLECULE KNOCKOUT MOUSE AND USES THEREOF
This invention relates to knockout mice for the Ca 2+ sensor membrane protein STIM-1, STIM-2, or both, as well as cell lines from these knockout mice. Provided herein are various methods of use of isolated with knockout STIM-1 and/or STIM-2.
1 . An isolated Stim-deficient cell, wherein the cell comprises a disruption of a STIM gene.
2 . The cell of claim 1 , wherein the cell is deficient in Stim1 protein, Stim2 protein, or both Stim1 and Stim2 proteins.
3 . The cell of claim 1 , wherein the disruption of the STIM gene is homozygous.
4 . The cell of claim 1 , wherein the disruption of the STIM gene is heterozygous.
5 . The cell of claim 1 , wherein the disruption of the STIM gene is conditional.
6 . The cell of claim 1 , wherein the disruption of the STIM gene is constitutive.
7 . The cell of claim 1 , wherein disruption of the STIM gene comprises a deletion, insertion, or substitution.
8 . The cell of claim 1 , wherein disruption of the STIM gene comprises recombination of the STIM gene.
9 . The cell of claim 1 , wherein the disruption of the STIM gene comprises a disruption of the coding sequence.
10 . A method of screening for agents that modulate intracellular calcium flux comprising contacting the isolated STIM-deficient cell of any of claims 1 - 9 with an agent.
11 . The method of claim 10 further comprising: (a) assessing the effects of the agent on Ca 2+ influx in the cell; and (b) identifying the agent as an agent that modulates intracellular Ca + if the agent has an effect on the intracellular Ca2+.
12 . A method for evaluating the mode of action of an agent that modulates intracellular calcium flux comprising contacting (i) the isolated STIM-deficient cell of any of claims 1 - 9 and (ii) a non STIM-deficient cell with an agent.
13 . The method of claim 12 further comprising: (a) assessing the effects of the agent on Ca 2+ influx in the non Stim-deficient cell and Stim-deficient cell; (b) comparing the effects of the agent on the Ca + influx in the non Stim-deficient cell and Stim-deficient cell; and (c) determining the agent as an agent that modulates intracellular Ca 2+ via a Stim-dependent pathway if the agent reduces the intracellular Ca + in the non Stim-deficient cell as compared to the Stim-deficient cell.
14 . A method for studying the cellular functions of a STIM protein or a mutant STIM protein in the absence of any other STIM homologues comprising introducing a STIM protein or a mutant STIM protein into the isolated STIM-deficient cell of any of claims 1 - 9 .
15 . The method of claim 14 , wherein the STIM protein or mutant STIM protein is STIM1 or STIM2.
16 . A method for screening a STIM inhibitor for side effects or toxicity resulting from the inhibitor's action on a target(s) other than STIM comprising contacting (i) the isolated STIM-deficient cell of any of claims 1 - 9 or (ii) a transgenic mouse whose genome comprises a heterozygous or homozygous disruption of a STIM gene with a STIM inhibitor.
17 . The method of claim 16 further comprising (a) monitoring the isolated STIM-deficient cell or the transgenic mouse to evaluate side effects or toxicity of the inhibitor.
18 . The method of claim 10 , wherein the agent is a biomolecule.
19 . The method of claim 18 , wherein the biomolecule is a nucleic acid or polypeptide.
20 . The method of claim 12 , wherein the agent is a biomolecule.