Methods for screening for transdominant effector peptides and RNA molecules

Abstract

Biochemical libraries are screened for transdominant intracellularly bioactive agents by expressing a molecular library of randomized nucleic acids as a plurality of corresponding expression products in a plurality of cells, each of the nucleic acids comprising a different nucleotide sequence, detecting a cell of the plurality of cells exhibiting a changed physiology in response to the presence in the cell of a transdominant expression product of the corresponding expressio products; and isolating the cell and/or transdominant expression product.

What is claimed is:  
     
         1 . A method of screening for a transdominant bioactive agent, said method comprising steps: 
 expressing a molecular library of randomized nucleic acids as a plurality of isolated corresponding randomized translation products in a first plurality of cells, each of said nucleic acids comprising a different nucleotide sequence;    screening a second plurality of cells for a cell exhibiting a changed physiology in response to the presence of a transdominant translation product of said plurality of isolated corresponding randomized translation products, wherein said translation product is expressed with a fusion partner, synthetic or heterologous to said first plurality of cells, comprising a localizing signal sequence capable of constitutively localizing said translation product to a predetermined subcellular locale, secretory and membrane-anchoring signal sequences capable of localizing said translation product to the plasma membrane, or a secretory signal sequence capable of effecting the secretion of said translation product.    detecting said cell;    isolating at least one of said cell and said transdominant translation product, wherein said transdominant translation product is a transdominant bioactive agent.    
     
     
         2 . A method according to  claim 1 , wherein said translation products are presented on the extracellular surface of said first plurality of cells.  
     
     
         3 . A method according to  claim 1 , wherein said translation products are secreted from said first plurality of cells.  
     
     
         4 . A method according to  claim 1 , wherein said first and second plurality of cells are different.  
     
     
         5 . A method according to  claim 1 , wherein said expressing step further comprises introducing said library into said cells.  
     
     
         6 . A method according to  claim 1 , wherein said expressing step further comprises introducing said library into said cells using retroviral vectors.  
     
     
         7 . A method of screening for a transdominant extracellularly bioactive agent, said method comprising steps: 
 expressing a molecular library of randomized nucleic acids as a plurality of isolated corresponding randomized translation products in a first plurality of cells, each of said nucleic acids comprising a different nucleotide sequence;    screening a second different plurality of cells for a cell exhibiting a changed physiology in response to the presence of a transdominant translation product of said plurality of isolated corresponding randomized translation products, wherein said translation product is expressed with a fusion partner, synthetic or heterologous to said first plurality of cells, comprising a secretory and membrane-anchoring signal sequences capable of localizing said translation product to the extracellular surface of the plasma membrane, or a secretory signal sequence capable of effecting the secretion of said translation product.    detecting said cell;    isolating at least one of said cell and said transdominant translation product, wherein said transdominant translation product is a transdominant extracellularly bioactive agent; wherein said expressing step comprises introducing said library into said cells using retroviral vectors.