Modifiable Chemical Inducers of Proximity and Methods of Using the Same

Abstract

Methods of reversibly inducing proximity of first and second target molecules in a sample are provided. Aspects of the methods include contacting the sample with a modifiable chemical inducer of proximity (MCIP) that reversibly induces proximity of the first and second target molecules, upon application of a stimulus that modifies the MCIP. Aspects of the invention further include methods for regulating a biological process in a cell. Aspects of the invention further include compositions, e.g., compounds and kits, etc., that find use in methods of the invention.

1 - 22 . (canceled) 
     
     
         23 . A modifiable chemical inducer of proximity (MCIP) described by formula (I): 
       
         
           
           
               
               
           
         
         wherein: 
         A is a first binding moiety that specifically binds a first target molecule; 
         B is a second binding moiety that specifically binds a second target molecule; and 
         X is a modifiable group that is modified in response to a stimulus. 
       
     
     
         24 . The MCIP of  claim 23 , wherein the first target molecule is a chimeric. 
     
     
         25 . The MCIP of  claim 23 , wherein the second target molecule is a chimeric. 
     
     
         26 . The MCIP of  claim 23 , wherein one or more of the first and second target molecules are chimeric proteins. 
     
     
         27 . The MCIP of  claim 23 , wherein the stimulus is a chemical agent. 
     
     
         28 . The MCIP of  claim 23 , wherein the stimulus is a photon. 
     
     
         29 . The MCIP of  claim 23 , wherein the stimulus is contact with an enzyme. 
     
     
         30 . The MCIP of  claim 23 , wherein the modifiable group is a cleavable group. 
     
     
         31 . The MCIP of  claim 30 , wherein the cleavable group is enzymatically cleavable. 
     
     
         32 . The MCIP of  claim 30 , wherein the cleavable group is photocleavable. 
     
     
         33 . The MCIP of  claim 30 , wherein X is part of A or B. 
     
     
         34 . The MCIP of  claim 33 , wherein A specifically binds a FKBP domain and B specifically binds a FRB domain. 
     
     
         35 . The MCIP of  claim 34 , wherein the MCIP is a rapamycin analog and X is a photoreactive group. 
     
     
         36 . The MCIP of  claim 35 , wherein X is a 7-nitroindolyl group located at the C7 position of the rapamycin analog. 
     
     
         37 . The MCIP of  claim 26 , wherein the MCIP is of formula (V):
   A-L-B   (V)
   wherein L is a linker that comprises X.   
     
     
         38 . The MCIP of  claim 37 , wherein A and B are homologous. 
     
     
         39 . The MCIP of  claim 37 , wherein A and B are heterologous. 
     
     
         40 . A kit comprising:
 an MCIP described by formula (I):   
       
         
           
           
               
               
           
         
         
           wherein: 
           A is a first binding moiety that specifically binds a first target domain; 
           B is a second binding moiety that specifically binds a second target domain; 
           X is a modifiable group that is modified in response to a stimulus; and 
         
         one or more components selected from:
 the first target domain, the second target domain, a first construct encoding the first target domain, a second construct encoding the second target domain, a cloning vector comprising a multiple cloning site (MSC), a cell, and a stimulus-applying component. 
 
       
     
     
         41 . The kit of  claim 40 , wherein one or more of the first and second target domains are part of a chimeric molecule. 
     
     
         42 . The kit of  claim 41 , wherein the first and second target domains are comprised in chimeric first and second target proteins. 
     
     
         43 . The kit of  claim 40 , wherein the first and second constructs are comprised in the same vector. 
     
     
         44 . The kit of  claim 40 , wherein the cloning vector expresses the first and second constructs via a bi-directional promoter or an internal ribosome entry site (IRES). 
     
     
         45 . The kit of  claim 40 , wherein, the MCIP comprises a cleavable rapamycin analog. 
     
     
         46 . The kit of  claim 40 , wherein the cell comprises the first and second constructs. 
     
     
         47 . The kit of  claim 40 , wherein the cell expresses the first and second target domains. 
     
     
         48 . The kit of  claim 40 , wherein the stimulus-applying component is a UV light source, an enzyme or a chemical agent.