Knock-in rodent comprising a mutation in an endogenous CRBN locus and methods of use thereof
The present invention features a knock-in mouse comprising a mutation in an endogenous CRBN locus and methods of use thereof.
1. A knock-in mouse whose genome comprises an exogenous polynucleotide encoding a mutant cereblon (CRBN) polypeptide at the endogenous CRBN locus, wherein said mutant CRBN comprises at least one S369C, V380E, or I391V substitution, and wherein the knock-in mouse has immunomodulatory imide drug (IMiD) sensitivity.
2. The knock-in mouse of claim 1 , wherein said mutant CRBN comprises an I391V substitution.
3. The knock-in mouse of claim 2 , wherein the mouse is pregnant.
4. A method of assessing the teratogenicity of an immunomodulatory imide drug (IMiD) in offspring of a pregnant mouse with IMiD sensitivity, the method comprising:
administering the IMiD to the pregnant mouse of claim 3 , and
assessing whether teratogenicity is induced in offspring of the pregnant mouse.
5. The method of claim 4 , wherein teratogenicity is assessed prenatally or postnatally.
6. A method of assessing immunomodulatory imide drug (IMiD) sensitivity of the transgenic mouse of claim 3 , the method comprising:
administering the IMiD to the mouse of claim 3 , and
assessing the amount of IMiD sensitivity in the mouse.
7. The method of claim 6 , wherein an increased amount of IMiD sensitivity is assessed by detecting one or more of the following:
an increased amount of IKZF1 or IKZF3 ubiquitination;
an increased amount of IKZF1 or IKZF3 degradation;
a decreased amount of IKZF1 or IKZF3 protein; and/or
an increased amount of binding of IKZF1 or IKZF3 to CRBN.
8. A method of assessing the ability of an agent to modulate immunomodulatory imide drug (IMiD) sensitivity using the knock-in mouse of claim 2 , the method comprising: administering the agent to the mouse of claim 2 , and assessing the amount of IMiD sensitivity in the mouse.
9. The method of claim 8 , wherein an increased amount of IMiD sensitivity is assessed by detecting one or more of the following:
an increased amount of IKZF1 or IKZF3 ubiquitination;
an increased amount of IKZF1 or IKZF3 degradation;
a decreased amount of IKZF1 or IKZF3 protein; and/or
an increased amount of binding of IKZF1 or IKZF3 to CRBN.
10. The knock-in mouse of claim 1 , wherein the IMiD is thalidomide, lenalidomide and/or pomalidomide.
11. A cell isolated from the mouse of claim 1 .
12. An isolated mouse cell whose genome comprises an exogenous polynucleotide encoding a mutant cereblon (CRBN) polypeptide at the endogenous CRBN locus, wherein said mutant CRBN comprises a I391V substitution capable of causing immunomodulatory imide drug (IMiD) sensitivity.
13. A method of assessing the ability of an agent to modulate immunomodulatory imide drug (IMiD) sensitivity using the isolated mouse cell of claim 12 , the method comprising: administering the agent to the isolated mouse cell of claim 12 , and assessing the amount of IMiD sensitivity in the mouse cell.
14. The method of claim 13 , wherein an increased amount of IMiD sensitivity is assessed by detecting one or more of the following:
an increased amount of IKZF1 or IKZF3 ubiquitination;
an increased amount of IKZF1 or IKZF3 degradation;
a decreased amount of IKZF1 or IKZF3 protein; and/or
an increased amount of binding of IKZF1 or IKZF3 to CRBN.