Staphylococcus aureus TargetArray: comprehensive differential essential gene expression as a mechanistic tool to profile antibacterials.
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.
Xu
H Howard
HH
Elitra Pharmaceuticals, San Diego, California 92121, USA. hxu3@calstatela.edu
Trawick
John D
JD
Haselbeck
Robert J
RJ
Forsyth
R Allyn
RA
Yamamoto
Robert T
RT
Archer
Rich
R
Patterson
Joe
J
Allen
Molly
M
Froelich
Jamie M
JM
Taylor
Ian
I
Nakaji
Danny
D
Maile
Randy
R
Kedar
G C
GC
Pilcher
Marshall
M
Brown-Driver
Vickie
V
McCarthy
Melissa
M
Files
Amy
A
Robbins
David
D
King
Paula
P
Sillaots
Susan
S
Malone
Cheryl
C
Zamudio
Carlos S
CS
Roemer
Terry
T
Wang
Liangsu
L
Youngman
Philip J
PJ
Wall
Daniel
D
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*
D000900:Anti-Bacterial Agents; D016372:RNA, Antisense