US 6,274,321 B1 · High throughput functional screening of cDNAs

Abstract

Methods for screening cDNAs that express a product interacting with a target molecule. Individual cDNAs are pooled and the cDNA pools are expressed to obtain expression products, for example by coupled in vitro transcription/translation. The interaction of the products with the target molecule is then assayed, for example by scintillation proximity assay (SPA), to identify pools of interest. By selectively re-pooling the cDNAs and repeating the expression and assay steps, individual cDNAs of interest can be rapidly identified. This method is readily automated in a computer-controlled device for high throughput screening. The invention also provides methods of transfecting a cell with a cDNA identified by the screening method to confer a desired property to a cell or identifying cDNAs from a pool of cDNAs by transfection into cells to confer a desired property.

Claims25

Dependent claims

A method of identifying a nucleic acid in a pool of interest, comprising the steps of

(1) obtaining a plurality of nucleic acids, wherein the nucleic acids are individually identifiable;
(2) pooling the nucleic acids in step (1) into at least two pools of at least one nucleic acid each;
(3) expressing the nucleic acid pools in step (2) to obtain corresponding protein expression product pools;
(4) assaying the expression product pools in step (3) for products having an interaction with a target molecule;
(5) selecting a nucleic acid pool corresponding to an expression product pool identified in step (4);
and
(6) identifying at least one individual nucleic acid in the nucleic acid pool identified in step (5).

4 more elements

Dependencies are inferred from the claim text.

High throughput functional screening of cDNAs

Life of this patent

High throughput functional screening of cDNAs

Life of this patent

Abstract

Claims25