Patent ID: 6830892
Filing Date: 2004-12-14
Classification: A61K,C07K,C12N,C40B

Abstract:
A method of screening for a target molecule from a group of target molecules comprising:(a) transducing a plurality of cells with a plurality of lentiviral virions, wherein the lentiviral virions are produced by: co-transfecting a producer cell with at least (1) one vector containing a lentiviral gag gene encoding a lentiviral gag protein, wherein the lentiviral gag gene is operably linked to a promoter and a polyadenylation sequence, (2) a second vector containing an env gene encoding a functional envelope protein, wherein the env gene is operably linked to a promoter and a polyadenylation sequence; and (3) a lentiviral pol gene encoding a lentiviral pol protein on one of the first two vectors or on at least a third vector, wherein said lentiviral pol gene is operably linked to a promoter and a polyadenylation sequence; wherein said vectors do not contain sufficient nucleotides to encode the lentiviral gag and pol and the envelope protein on a single vector; and wherein said vectors do not contain nucleotides of a lentiviral genome referred to as a packaging segment to effectively package lentiviral RNA; and wherein the lentiviral proteins and the envelope protein when expressed in combination form a lentivirus virion containing an envelope protein around a lentiviral capsid; and (4) a packaging vector containing a nucleic acid sequence encoding a target molecule selected from a plurality of target molecules, wherein the nucleic acid sequence is operably linked to a promoter and a lentiviral packaging sequence including the portion of the lentiviral long terminal repeat (LTR) sequences necessary to package the lentiviral RNA into the lentiviral virion; wherein the virion further contains a marker gene; (b) identifying transduced cells by screening for the presence of a marker expressed by the marker gene; (c) screening for a cell displaying a desired phenotype; and (d) identifying the target molecule present in the cell displaying the desired phenotype.