ZFIN ID: ZDB-PUB-090424-2
Nitroreductase-mediated cell ablation in transgenic zebrafish embryos
Pisharath, H., and Parsons, M.J.
Date: 2009
Source: Methods in molecular biology (Clifton, N.J.)   546: 133-143 (Chapter)
Registered Authors: Parsons, Michael
Keywords: Ablation, Nitroreductase, Metronidazole, Prodrug, Transactivation, Gal4, UAS
MeSH Terms:
  • Animals
  • Animals, Genetically Modified/embryology
  • Animals, Genetically Modified/genetics
  • Animals, Genetically Modified/metabolism*
  • Anti-Infective Agents/pharmacology
  • Apoptosis/drug effects*
  • DNA/genetics
  • DNA/metabolism
  • Escherichia coli Proteins/administration & dosage*
  • Escherichia coli Proteins/biosynthesis
  • Genetic Engineering/methods
  • Green Fluorescent Proteins
  • Metronidazole/pharmacology
  • Microinjections
  • Microscopy, Confocal
  • Nitroreductases/administration & dosage*
  • Nitroreductases/biosynthesis
  • Prodrugs/pharmacology
  • Recombinant Fusion Proteins/administration & dosage
  • Recombinant Fusion Proteins/biosynthesis
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish/metabolism*
PubMed: 19378102 Full text @ Meth. Mol. Biol.
Prodrug dependent cell ablation is a method that allows inducible and spatially restricted cell destruction. We describe transgenic methods to express the Escherichia coli nfsB in a tissue restricted manner in the zebrafish. This bacterial gene encodes a nitroreductase (NTR) enzyme that can render prodrugs such as metronidazole (Met) cytotoxic. Using the expression of NTR fused to a fluorescent protein, one can simultaneously make cells susceptible to prodrug treatment and visualize cell ablation as it occurs.