Methods and compositions for detection of targets involved in cancer metastasis

Abstract

The present invention is further directed to methods and compositions for identifying target nucleic acids that are involved in cancer metastasis.

1 . A method of identifying one or more genes that modulate tumor metastasis, the method comprising:
 (a) transfecting an immortalized cell line with one or more target cDNAs to produce transfected cells, wherein the transfected cells further express a detectable reporter gene, and wherein the immortalized cell line lacks metastatic potential in the absence of the transfected target cDNAs;   (b) injecting the transfected cells into a non-human mammal;   (c) imaging the injected animal model to detect tumor metastasis;   (d) isolating tumor cells from the injected animal model;   (e) analyzing the isolated tumor cells to identify whether open reading frames from the one or more target cDNAs are integrated into genomes of the tumor cells, thereby identifying one or more genes that modulate tumor metastasis.   
     
     
         2 . The method of  claim 1 , wherein the animal model is a mouse. 
     
     
         3 . The method of  claim 1 , wherein the animal model is an immune-deficient mouse that lacks B- and T-cell lineages. 
     
     
         4 . The method of  claim 1 , wherein the one or more target cDNAs comprise a library of at least 100 cDNAs. 
     
     
         5 . The method of  claim 1 , wherein the one or more target cDNAs comprise genes encoding transmembrane proteins. 
     
     
         6 . The method of  claim 1 , wherein prior to isolating step (d), the injected animal model is imaged periodically for at least 50 days. 
     
     
         7 . The method of  claim 1 , wherein the one or more target cDNAs comprises at least 80 pools of genes, wherein each pool comprises at least 40 genes. 
     
     
         8 . The method of  claim 1 , wherein different cells are transfected with different cDNAs, such that each transfected cell expresses a single target cDNA. 
     
     
         9 . The method of  claim 1 , wherein the analyzing is conducted using a method selected from the group consisting of PCR, Southern Blot, and qRT-PCR. 
     
     
         10 . The method of  claim 1 , wherein each of the target cDNAs comprises an open reading frame of a gene encoding a transmembrane receptor and a CMV promoter. 
     
     
         11 . The method of  claim 1 , wherein the detectable reporter gene encodes a luciferase.