NTR 2.0: A rationally-engineered prodrug converting enzyme with substantially enhanced efficacy for targeted cell ablation

Abigail V. Sharrock, Timothy S. Mulligan, Kelsi R. Hall, Elsie M. Williams, David T. White, Liyun Zhang, Frazer Matthews, Saumya Nimmagadda, Selena Washington, Katherine Le, Danielle Meir-Levi, Olivia L. Cox, Meera T. Saxena, Anne L. Calof, Martha E. Lopez-Burks, Arthur D. Lander, Ding Ding, Hongkai Ji, David F. Ackerley, Jeffrey Mumm

Research output: Contribution to journalArticlepeer-review

Abstract

Heterologously-expressed bacterial nitroreductase (NTR) enzymes sensitize eukaryotic cells to prodrugs such as metronidazole (MTZ), enabling selective cell ablation paradigms that have expanded studies of cell function and regeneration in vertebrate systems. However, first-generation NTRs require confoundingly toxic prodrug treatments (e.g. 10 mM MTZ) and some cell types have proven resistant. We used rational engineering and cross-species screening to develop a NTR variant, NTR 2.0, which exhibits ~100-fold improvement in MTZ-mediated cell-specific ablation efficacy. Toxicity tests in zebrafish showed no deleterious effects of prolonged MTZ treatments of ≤1 mM. NTR 2.0 therefore enables sustained cell loss paradigms and ablation of previously resistant cell types. These properties permit enhanced interrogations of cell function, extended challenges to the regenerative capacities of discrete stem cell niches, and enable modeling of chronic degenerative diseases. Accordingly, we have created a series of bipartite transgenic resources to facilitate dissemination of NTR 2.0 to the research community.

Original languageEnglish (US)
JournalUnknown Journal
DOIs
StatePublished - May 24 2020

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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