title: A genome-wide strategy for the identification of essential genes in Staphylococcus aureus.
abstract: To address the need for new approaches to antibiotic drug development, we have identified a large number of essential genes for the bacterial pathogen, Staphylococcus aureus, using a rapid shotgun antisense RNA method. Staphylococcus aureus chromosomal DNA fragments were cloned into a xylose-inducible expression plasmid and transformed into S. aureus. Homology comparisons between 658 S. aureus genes identified in this particular antisense screen and the Mycoplasma genitalium genome, which contains 517 genes in total, yielded 168 conserved genes, many of which appear to be essential in M. genitalium and other bacteria. Examples are presented in which expression of an antisense RNA specifically reduces its cognate mRNA. A cell-based, drug-screening assay is also described, wherein expression of an antisense RNA confers specific sensitivity to compounds targeting that gene product. This approach enables facile assay development for high throughput screening for any essential gene, independent of its biochemical function, thereby greatly facilitating the search for new antibiotics.
authors.affiliation: Elitra Pharmaceuticals, San Diego, CA 92121, USA.
authors.forename: R Allyn
authors.initials: RA
authors.lastname: Forsyth
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Haselbeck
authors.forename: Kari L
authors.initials: KL
authors.lastname: Ohlsen
authors.forename: Robert T
authors.initials: RT
authors.lastname: Yamamoto
authors.forename: Howard
authors.initials: H
authors.lastname: Xu
authors.forename: John D
authors.initials: JD
authors.lastname: Trawick
authors.forename: Daniel
authors.initials: D
authors.lastname: Wall
authors.forename: Liangsu
authors.initials: L
authors.lastname: Wang
authors.forename: Vickie
authors.initials: V
authors.lastname: Brown-Driver
authors.forename: Jamie M
authors.initials: JM
authors.lastname: Froelich
authors.forename: Kedar G
authors.initials: KG
authors.lastname: C
authors.forename: Paula
authors.initials: P
authors.lastname: King
authors.forename: Melissa
authors.initials: M
authors.lastname: McCarthy
authors.forename: Cheryl
authors.initials: C
authors.lastname: Malone
authors.forename: Brian
authors.initials: B
authors.lastname: Misiner
authors.forename: David
authors.initials: D
authors.lastname: Robbins
authors.forename: Zehui
authors.initials: Z
authors.lastname: Tan
authors.forename: Zhan-yang
authors.initials: ZY
authors.lastname: Zhu Zy
authors.forename: Grant
authors.initials: G
authors.lastname: Carr
authors.forename: Deborah A
authors.initials: DA
authors.lastname: Mosca
authors.forename: Carlos
authors.initials: C
authors.lastname: Zamudio
authors.forename: J Gordon
authors.initials: JG
authors.lastname: Foulkes
authors.forename: Judith W
authors.initials: JW
authors.lastname: Zyskind
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D019295:Computational Biology / Q000379:methods; D018390:Gene Targeting; D020043:Genes, Essential; D016680:Genome, Bacterial; D009174:Mycoplasma / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D016372:RNA, Antisense; D012333:RNA, Messenger / Q000235:genetics; D013211:Staphylococcus aureus / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D014169:Transformation, Bacterial; D014994:Xylose / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D016372:RNA, Antisense; D012333:RNA, Messenger; D014994:Xylose