Abstract
There are two main strategies for antibiotic discovery: target-based and phenotypic screening. The latter has been much more successful in delivering first-in-class antibiotics, despite the major bottleneck of delayed Mechanism-of-Action (MOA) identification. Although finding new antimicrobial compounds is a very challenging task, identifying their MOA has proven equally challenging. MOA identification is important because it is a great facilitator of lead optimization and improves the chances of commercialization. Moreover, the ability to rapidly detect MOA could enable a shift from an activity-based discovery paradigm towards a mechanism-based approach. This would allow to probe the grey chemical matter, an underexplored source of structural novelty. In this study we review techniques with throughput suitable to screen large libraries and sufficient sensitivity to distinguish MOA. In particular, the techniques used in chemical genetics (e.g., based on overexpression and knockout/knockdown collections), promoter-reporter libraries, transcriptomics (e.g., using microarrays and RNA sequencing), proteomics (e.g., either gel-based or gel-free techniques), metabolomics (e.g., resourcing to nuclear magnetic resonance or mass spectrometry techniques), bacterial cytological profiling, and vibrational spectroscopy (e.g., Fourier-transform infrared or Raman scattering spectroscopy) were discussed. Ultimately, new and reinvigorated phenotypic assays bring renewed hope in the discovery of a new generation of antibiotics.
Highlights
There are two main strategies for antibiotic discovery: target-based and phenotypic screening
This results in higher rates of rediscovery, which is a key challenge in natural product antibiotic discovery [5] and an inability to detect low potency candidates, which can be later modified for enhanced therapeutic efprocess fect
[6]. and require subsequent efforts [4]. This results in higher rates of rediscovery, which is a key challenge in natural product antibiotic discoveryis[5]
Summary
Chemical genetics evaluates a library of chemical compounds against a. Chemical evaluates of chemical compounds genome library, i.e., itgenetics maps the effect ofa alibrary wide set of exogenous ligands against across aa genome wide library, i.e., it maps the effect of a wide set of exogenous ligands across a wide wide set of genetic variants of cells models [18] (Figure 2A). When the exogenous ligand is ligand is an antibiotic candidate, the effect across a mutant library enlightens its MOA. High-throughput screening assays basedthe ontotal genome-wide or promoter-wide mutant libraries that are against a chemical library. The total number of genes (g) or promoters (p) considered in a screening campaign are minimized to reduce the workload, this often yields less information.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
