title: Creation of genome-wide protein expression libraries using random activation of gene expression.
abstract: Here we report the use of random activation of gene expression (RAGE) to create genome-wide protein expression libraries. RAGE libraries containing only 5 x 10(6) individual clones were found to express every gene tested, including genes that are normally silent in the parent cell line. Furthermore, endogenous genes were activated at similar frequencies and expressed at similar levels within RAGE libraries created from multiple human cell lines, demonstrating that RAGE libraries are inherently normalized. Pools of RAGE clones were used to isolate 19,547 human gene clusters, approximately 53% of which were novel when tested against public databases of expressed sequence tag (EST) and complementary DNA (cDNA). Isolation of individual clones confirmed that the activated endogenous genes can be expressed at high levels to produce biologically active proteins. The properties of RAGE libraries and RAGE expression clones are well suited for a number of biotechnological applications including gene discovery, protein characterization, drug development, and protein manufacturing.
authors.affiliation: Athersys, Inc., 3201 Carnegie Ave., Cleveland, OH 44115, USA. jharrington@athersys.com
authors.forename: J J
authors.initials: JJ
authors.lastname: Harrington
authors.forename: B
authors.initials: B
authors.lastname: Sherf
authors.forename: S
authors.initials: S
authors.lastname: Rundlett
authors.forename: P D
authors.initials: PD
authors.lastname: Jackson
authors.forename: R
authors.initials: R
authors.lastname: Perry
authors.forename: S
authors.initials: S
authors.lastname: Cain
authors.forename: C
authors.initials: C
authors.lastname: Leventhal
authors.forename: M
authors.initials: M
authors.lastname: Thornton
authors.forename: R
authors.initials: R
authors.lastname: Ramachandran
authors.forename: J
authors.initials: J
authors.lastname: Whittington
authors.forename: L
authors.initials: L
authors.lastname: Lerner
authors.forename: D
authors.initials: D
authors.lastname: Costanzo
authors.forename: K
authors.initials: K
authors.lastname: McElligott
authors.forename: S
authors.initials: S
authors.lastname: Boozer
authors.forename: R
authors.initials: R
authors.lastname: Mays
authors.forename: E
authors.initials: E
authors.lastname: Smith
authors.forename: N
authors.initials: N
authors.lastname: Veloso
authors.forename: A
authors.initials: A
authors.lastname: Klika
authors.forename: J
authors.initials: J
authors.lastname: Hess
authors.forename: K
authors.initials: K
authors.lastname: Cothren
authors.forename: K
authors.initials: K
authors.lastname: Lo
authors.forename: J
authors.initials: J
authors.lastname: Offenbacher
authors.forename: J
authors.initials: J
authors.lastname: Danzig
authors.forename: M
authors.initials: M
authors.lastname: Ducar
mesh_terms: D002460:Cell Line; D016208:Databases, Factual; D004797:Enzyme-Linked Immunosorbent Assay; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D005787:Gene Frequency; D005821:Genetic Techniques; D015894:Genome, Human; D015698:Genomic Library; D006801:Humans; D008969:Molecular Sequence Data; D005810:Multigene Family; D011506:Proteins / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016324:Sequence Tagged Sites
chemical_list: D011506:Proteins