Nitroreductase-mediated cell ablation in transgenic zebrafish embryos.
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.
Pisharath
Harshan
H
Department of Surgery, Johns Hopkins School of Medicine, Baltimore, MD, 21205, USA.
Parsons
Michael J
MJ
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology / Q000235:genetics / Q000378:metabolism*; D000890:Anti-Infective Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000008:administration & dosage* / Q000096:biosynthesis; D005818:Genetic Engineering / Q000379:methods; D049452:Green Fluorescent Proteins; D008795:Metronidazole / Q000494:pharmacology; D008845:Microinjections; D018613:Microscopy, Confocal; D009601:Nitroreductases / Q000008:administration & dosage* / Q000096:biosynthesis; D011355:Prodrugs / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000096:biosynthesis; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism*
D000890:Anti-Infective Agents; D029968:Escherichia coli Proteins; D011355:Prodrugs; D011993:Recombinant Fusion Proteins; D008795:Metronidazole; D049452:Green Fluorescent Proteins; D004247:DNA; C116054:NfsB protein, E coli; D009601:Nitroreductases