Patent ID: 7427474

Claim:
A method of identifying a function of a polypeptide-encoding sequence of interest endogenously expressed by a cell type using high throughput detection, comprising: a) providing a first pseudotyped lentiviral vector, comprising at least a part of the polypeptide-encoding sequence of interest, wherein the polypeptide-encoding sequence of interest is a known or unidentified gene sequence, and wherein the first lentiviral vector is designed to express little or no vector borne sequence other than the at least a part of the polypeptide-encoding sequence of interest, and b) providing a second pseudotyped lentiviral vector, comprising an inhibitory or termination sequence, wherein the inhibitory or termination sequence can inhibit, terminate, or underexpress the endogenously expressed polypeptide-encoding sequence of interest, and wherein the second lentiviral vector is designed to express little or no vector borne sequence other than the inhibitory or termination sequence, c) providing a first and a second population of the cell type, and transducing the first lentiviral vector in the first cell population and transducing the second lentiviral vector in the second cell population; d) expressing all or part of the polypeptide-encoding sequence of interest in the first cell population and inhibiting, terminating, or underexpressing the polypeptide-encoding sequence of interest in the second cell population; e) high throughput detecting at least one change in one or more endogenous cellular factors in the first and second cell populations by comparing the effect on the cell of expression of the polypeptide-encoding sequence of interest with the effect on the cell of inhibition, or termination, or underexpression of the polypeptide-encoding sequence of interest; and f) identifying a function of the polypeptide-encoding sequence of interest based on the detected and compared effect on the cell of expression and inhibition, or termination, or underexpression of the polypeptide-encoding sequence of interest on one or more cellular factors.