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Selective Cell Ablation Using an Improved Prodrug-Converting Nitroreductase.

Timothy S Mulligan1, Jeff S Mumm2

  • 1Department of Ophthalmology, Wilmer Eye Institute, Johns Hopkins University, Baltimore, MD, USA.

Methods in Molecular Biology (Clifton, N.J.)
|September 5, 2023
PubMed
Summary
This summary is machine-generated.

A new nitroreductase (NTR) variant, NTR 2.0, enables precise cell ablation in zebrafish. This improved system allows targeted cell elimination at lower drug doses, aiding regeneration studies and disease modeling.

Keywords:
AblationCell ablationMTZMetronidazoleNTRNTR 1.1NTR 2.0NfsBNifurpirinolNitroreductaseProdrugRonidazoleZebrafish

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Area of Science:

  • Developmental Biology
  • Regenerative Medicine
  • Toxicology

Background:

  • Selective cell ablation is crucial for studying cell function, regeneration, and disease modeling.
  • The nitroreductase (NTR)-mediated system uses an enzyme and prodrug (metronidazole) for targeted cell death.
  • Existing methods face limitations with certain cell types and potential toxicity.

Purpose of the Study:

  • To introduce an improved nitroreductase enzyme (NTR 2.0) for enhanced targeted cell ablation in zebrafish.
  • To enable spatially and temporally controlled cell elimination with reduced side effects.
  • To expand the utility of cell ablation for investigating regeneration and disease.

Main Methods:

  • Development and application of a novel nitroreductase enzyme variant (NTR 2.0).
  • Utilizing metronidazole as a prodrug for NTR-mediated cell death induction.
  • Employing transgenic techniques for controlled enzyme expression in zebrafish.

Main Results:

  • NTR 2.0 achieves effective cell ablation at significantly lower metronidazole concentrations.
  • The system demonstrates efficacy in ablating previously resistant cell types.
  • Reduced prodrug concentrations minimize off-target toxic effects.

Conclusions:

  • NTR 2.0 offers a more precise and safer method for targeted cell ablation in zebrafish.
  • This advancement facilitates deeper understanding of cell function, regeneration, and chronic inflammatory/degenerative diseases.
  • The improved system opens new avenues for regenerative medicine research and disease modeling.