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Related Concept Videos

Rapid Identification of Pathogens01:25

Rapid Identification of Pathogens

MALDI-TOF MS has transformed clinical microbiology by offering a rapid and reliable method for pathogen identification. The traditional approach to microbial identification typically involves time-consuming culture techniques and biochemical tests, which can delay the initiation of appropriate antimicrobial therapy. MALDI-TOF MS avoids these delays by using characteristic ribosomal protein mass patterns of microbial cells, enabling accurate species-level identification within minutes.Principle...

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Related Experiment Video

Updated: Jun 16, 2026

A Microscopic Phenotypic Assay for the Quantification of Intracellular Mycobacteria Adapted for High-throughput/High-content Screening
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A Multiplexed, Target-Based Phenotypic Screening Platform Using CRISPR Interference in Mycobacterium abscessus.

Donavan Marcus Neo1,2,3, Ishay Ben-Zion1, Josephine Bagnall1

  • 1Center for Integrated Solutions for Infectious Diseases, Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, United States.

ACS Infectious Diseases
|October 27, 2025
PubMed
Summary
This summary is machine-generated.

New PROSPECT (PRimary screening Of Strains to Prioritize Expanded Chemistry and Targets) strategy uses CRISPR interference to find new drugs for difficult-to-treat Mycobacterium abscessus infections, identifying compounds targeting InhA.

Keywords:
CRISPR interferenceMycobacterium abscessusantibiotic discoverychemical–genetic interactionscompound screeninghigh-throughput multiplexed screeningisoniazid intrinsic resistance

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Area of Science:

  • Microbiology
  • Drug Discovery
  • Genetics

Background:

  • Mycobacterium abscessus infections are difficult to treat due to extensive antibiotic resistance.
  • Limited success in discovering novel antimicrobial compounds necessitates innovative drug discovery strategies.

Purpose of the Study:

  • To advance the PROSPECT (PRimary screening Of Strains to Prioritize Expanded Chemistry and Targets) antimicrobial discovery strategy for Mycobacterium abscessus.
  • To leverage CRISPR interference (CRISPRi) for efficient generation of essential gene-depleted mutants.

Main Methods:

  • Modified PROSPECT strategy using CRISPRi to create pooled essential gene-depleted mutants in M. abscessus.
  • Screened a library of 782 compounds using CRISPRi guides as mutant barcodes.
  • Identified chemical-genetic interactions to find whole-cell active compounds.

Main Results:

  • Identified active compounds, including those targeting InhA, a key mycobacterial enzyme.
  • Discovered unexpected susceptibility to isoniazid, suggesting complex resistance mechanisms.
  • Demonstrated the utility of PROSPECT with CRISPRi for M. abscessus drug discovery.

Conclusions:

  • PROSPECT combined with CRISPRi engineering offers an accessible, high-throughput screening platform.
  • This approach accelerates early-stage drug discovery for challenging M. abscessus infections.
  • Further research is needed to develop new therapeutic options for M. abscessus.