Abstract
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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Animals, Genetically Modified / embryology
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Animals, Genetically Modified / genetics
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Animals, Genetically Modified / metabolism*
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Anti-Infective Agents / pharmacology
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Apoptosis / drug effects*
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DNA / genetics
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DNA / metabolism
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Escherichia coli Proteins / administration & dosage*
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Escherichia coli Proteins / biosynthesis
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Genetic Engineering / methods
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Green Fluorescent Proteins
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Metronidazole / pharmacology
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Microinjections
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Microscopy, Confocal
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Nitroreductases / administration & dosage*
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Nitroreductases / biosynthesis
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Prodrugs / pharmacology
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Recombinant Fusion Proteins / administration & dosage
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Recombinant Fusion Proteins / biosynthesis
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Zebrafish / embryology
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Zebrafish / genetics*
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Zebrafish / metabolism*
Substances
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Anti-Infective Agents
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Escherichia coli Proteins
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Prodrugs
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Recombinant Fusion Proteins
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Metronidazole
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Green Fluorescent Proteins
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DNA
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NfsB protein, E coli
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Nitroreductases