Rapid Gene Silencing Followed by Antimicrobial Susceptibility Testing for Target Validation in Antibiotic Discovery.

Methods Mol Biol

Mycobacteria Research Laboratory, Institute of Structural and Molecular Biology, School of Natural Sciences, Birkbeck, University of London, London, UK.

Published: July 2024

Mycobacterium tuberculosis is the main causative agent of tuberculosis (TB)-an ancient yet widespread global infectious disease to which 1.6 million people lost their lives in 2021. Antimicrobial resistance (AMR) has been an ongoing crisis for decades; 4.95 million deaths were associated with antibiotic resistance in 2019. While AMR is a multi-faceted problem, drug discovery is an urgent part of the solution and is at the forefront of modern research.The landscape of drug discovery for TB has undoubtedly been transformed by the development of high-throughput gene-silencing techniques that enable interrogation of every gene in the genome, and their relative contribution to fitness, virulence, and AMR. A recent advance in this area is CRISPR interference (CRISPRi). The application of this technique to antimicrobial susceptibility testing (AST) is the subject of ongoing research in basic science.CRISPRi technology can be used in conjunction with the high-throughput SPOT-culture growth inhibition assay (HT-SPOTi) to rapidly evaluate and assess gene essentiality including non-essential, conditionally essential (by using appropriate culture conditions), and essential genes. In addition, the HT-SPOTi method can develop drug susceptibility and drug resistance profiles.This technology is further useful for drug discovery groups who have designed target-based inhibitors rationally and wish to validate the primary mechanisms of their novel compounds' antibiotic action against the proposed target.

Download full-text PDF

Source
http://dx.doi.org/10.1007/978-1-0716-3981-8_3DOI Listing

Publication Analysis

Top Keywords

drug discovery
12
antimicrobial susceptibility
8
susceptibility testing
8
drug
5
rapid gene
4
gene silencing
4
silencing antimicrobial
4
testing target
4
target validation
4
validation antibiotic
4

Similar Publications

Discovery of noncovalent diaminopyrimidine-based Inhibitors for glioblastoma via a dual FAK/DNA targeting strategy.

Eur J Med Chem

January 2025

School of Pharmaceutical Sciences, Guizhou University, Guiyang, 550025, China. Electronic address:

Temozolomide, a widely used alkylating agent for glioblastoma treatment, faces significant challenges due to the development of resistance, which severely impacts patient survival. This underscores the urgent need for novel strategies to overcome this barrier. Focal adhesion kinase (FAK), an intracellular non-receptor tyrosine kinase, is highly expressed in glioblastoma cells and has been identified as a promising therapeutic target for anti-glioblastoma drug development.

View Article and Find Full Text PDF

Anxiolytic-like Effect of Chrysin on Female Zebrafish is Likely Mediated by α5 subunits of GABAA Receptors.

Chem Biodivers

January 2025

UNIFESSPA: Universidade Federal do Sul e Sudeste do Para, Faculdade de Psicologia, Rod. BR-230 (Transamazônica), Loteamento Cidade Jardim, Av. dos Ipês, s/n.º - Ci, 68503000, Marabá, BRAZIL.

Chrysin (5,7-dihydroxyflavone) is a natural flavonoid with potential anxiolytic-like effects in preclinical models. Acute treatment with this molecule (0 - 10 mg/kg) produced a biphasic dose-response in the zebrafish light/dark test (LDT), with anxiolytic-like effect at low doses and anxiogenic-like effects at high doses. Chrysin (1 mg/kg) decreased anxiety-like behavior in the zebrafish novel tank test (NTT), but did not prevent the anxiogenic effects of acute stress.

View Article and Find Full Text PDF

Repurposing the familiar: Future treatment options against chronic kidney disease.

J Pharm Pharmacol

January 2025

Department of Pharmacy, Birla Institute of Technology and Science, Pilani, Pilani Campus, 333031, Rajasthan, India.

Objectives: Chronic kidney disease (CKD) is a serious health issue with rising morbidity and mortality rates. Despite advances in understanding its pathophysiology, effective therapeutic options are limited, necessitating innovative treatment approaches. Also, current frontline treatments that are available against CKD are not uniformly effective and often come with significant side effects.

View Article and Find Full Text PDF

More than 470 million people globally are infected with the hookworms Ancylostoma ceylanicum and Necator americanus, resulting in an annual loss of 2.1 to 4 million disability-adjusted-life-years. Current infection management approaches are limited by modest drug efficacy, the costs associated with frequent mass drug administration campaigns, and the risk of reinfection and burgeoning drug resistance.

View Article and Find Full Text PDF

PEGylation of Dipeptide Linker Improves Therapeutic Index and Pharmacokinetics of Antibody-Drug Conjugates.

Bioconjug Chem

January 2025

Biotherapeutics Discovery Research Center, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201203, China.

Hydrophobic payloads incorporated into antibody-drug conjugates (ADCs) typically are superior to hydrophilic ones in tumor penetration and "bystander killing" upon release from ADCs. However, they are prone to aggregation and accelerated plasma clearance, which lead to reduced efficacies and increased toxicities of ADC molecules. Shielding the hydrophobicity of payloads by incorporating polyethylene glycol (PEG) elements or sugar groups into the ADC linkers has emerged as a viable alternative to directly adopting hydrophilic payloads.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!