Bulletin of the American Physical Society
APS March Meeting 2023
Volume 68, Number 3
Las Vegas, Nevada (March 5-10)
Virtual (March 20-22); Time Zone: Pacific Time
Session G10: Quantitative Cell Physiology II - Metabolism and Growth
11:30 AM–2:30 PM,
Tuesday, March 7, 2023
Room: Room 202
Sponsoring
Unit:
DBIO
Chair: Terence Hwa, University of California, San Diego
Abstract: G10.00014 : Utilizing massively parallel CRISPRi assays to investigate antibiotic tolerance*
2:06 PM–2:18 PM
Presenter:
Keiran Stevenson
(Cornell University)
Authors:
Keiran Stevenson
(Cornell University)
Guillaume Lambert
(Cornell University)
Louis B Cortes
(Cornell University)
We demonstrate that this approach can be used to investigate antibiotic tolerance and persisters by controlling the cas protein using an inducible promoter and thereby allowing for the reversal of growth based effects.
In addition the mechanism can be easily coupled with fluorescence markers and single gene targets can be made allowing for microscopy investigation of the physiological effects of th e cells.
We further use our technique to explore antibiotic tolerance in Escherichia coli on exposure to Naladixic acid, Carbenicillin and Tetracycline and highlight several gene categories that have gone otherwise unrecognised as being important in antibiotic survival. We further use single cell microscopy using microfluidics to attempt to illuminate the difference between different targets and their recovery using our knockdown system.
*This work was supported by NIH funding under 668 1R35 GM133759 Maximizing Investigators’ Research Award (MIRA)
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