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.
Department of Surgery, Johns Hopkins School of Medicine, Baltimore, MD, 21205, USA.
Harshan
H
Pisharath
Michael J
MJ
Parsons
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