title: Staphylococcus aureus TargetArray: comprehensive differential essential gene expression as a mechanistic tool to profile antibacterials.
abstract: The widespread emergence of antibiotic-resistant bacteria and a lack of new pharmaceutical development have catalyzed a need for new and innovative approaches for antibiotic drug discovery. One bottleneck in antibiotic discovery is the lack of a rapid and comprehensive method to identify compound mode of action (MOA). Since a hallmark of antibiotic action is as an inhibitor of essential cellular targets and processes, we identify a set of 308 essential genes in the clinically important pathogen Staphylococcus aureus. A total of 446 strains differentially expressing these genes were constructed in a comprehensive platform of sensitized and resistant strains. A subset of strains allows either target underexpression or target overexpression by heterologous promoter replacements with a suite of tetracycline-regulatable promoters. A further subset of 236 antisense RNA-expressing clones allows knockdown expression of cognate targets. Knockdown expression confers selective antibiotic hypersensitivity, while target overexpression confers resistance. The antisense strains were configured into a TargetArray in which pools of sensitized strains were challenged in fitness tests. A rapid detection method measures strain responses toward antibiotics. The TargetArray antibiotic fitness test results show mechanistically informative biological fingerprints that allow MOA elucidation.
authors.affiliation: Elitra Pharmaceuticals, San Diego, California 92121, USA. hxu3@calstatela.edu
authors.forename: H Howard
authors.initials: HH
authors.lastname: Xu
authors.forename: John D
authors.initials: JD
authors.lastname: Trawick
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Haselbeck
authors.forename: R Allyn
authors.initials: RA
authors.lastname: Forsyth
authors.forename: Robert T
authors.initials: RT
authors.lastname: Yamamoto
authors.forename: Rich
authors.initials: R
authors.lastname: Archer
authors.forename: Joe
authors.initials: J
authors.lastname: Patterson
authors.forename: Molly
authors.initials: M
authors.lastname: Allen
authors.forename: Jamie M
authors.initials: JM
authors.lastname: Froelich
authors.forename: Ian
authors.initials: I
authors.lastname: Taylor
authors.forename: Danny
authors.initials: D
authors.lastname: Nakaji
authors.forename: Randy
authors.initials: R
authors.lastname: Maile
authors.forename: G C
authors.initials: GC
authors.lastname: Kedar
authors.forename: Marshall
authors.initials: M
authors.lastname: Pilcher
authors.forename: Vickie
authors.initials: V
authors.lastname: Brown-Driver
authors.forename: Melissa
authors.initials: M
authors.lastname: McCarthy
authors.forename: Amy
authors.initials: A
authors.lastname: Files
authors.forename: David
authors.initials: D
authors.lastname: Robbins
authors.forename: Paula
authors.initials: P
authors.lastname: King
authors.forename: Susan
authors.initials: S
authors.lastname: Sillaots
authors.forename: Cheryl
authors.initials: C
authors.lastname: Malone
authors.forename: Carlos S
authors.initials: CS
authors.lastname: Zamudio
authors.forename: Terry
authors.initials: T
authors.lastname: Roemer
authors.forename: Liangsu
authors.initials: L
authors.lastname: Wang
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Youngman
authors.forename: Daniel
authors.initials: D
authors.lastname: Wall
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000235:genetics*; D020043:Genes, Essential / Q000235:genetics*; D016372:RNA, Antisense / Q000235:genetics; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics*
chemical_list: D000900:Anti-Bacterial Agents; D016372:RNA, Antisense