NTR 2.0: a rationally engineered prodrug-converting enzyme with substantially enhanced efficacy for targeted cell ablation.
Publication/Presentation Date
2-1-2022
Abstract
Transgenic expression of bacterial nitroreductase (NTR) enzymes sensitizes eukaryotic cells to prodrugs such as metronidazole (MTZ), enabling selective cell-ablation paradigms that have expanded studies of cell function and regeneration in vertebrates. However, first-generation NTRs required confoundingly toxic prodrug treatments to achieve effective cell ablation, and some cell types have proven resistant. Here we used rational engineering and cross-species screening to develop an NTR variant, NTR 2.0, which exhibits ~100-fold improvement in MTZ-mediated cell-specific ablation efficacy, eliminating the need for near-toxic prodrug treatment regimens. 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 novel modeling of chronic degenerative diseases. Accordingly, we have created a series of bipartite transgenic reporter/effector resources to facilitate dissemination of NTR 2.0 to the research community.
Volume
19
Issue
2
First Page
205
Last Page
215
ISSN
1548-7105
Published In/Presented At
Sharrock, A. V., Mulligan, T. S., Hall, K. R., Williams, E. M., White, D. T., Zhang, L., Emmerich, K., Matthews, F., Nimmagadda, S., Washington, S., Le, K. D., Meir-Levi, D., Cox, O. L., Saxena, M. T., Calof, A. L., Lopez-Burks, M. E., Lander, A. D., Ding, D., Ji, H., Ackerley, D. F., … Mumm, J. S. (2022). NTR 2.0: a rationally engineered prodrug-converting enzyme with substantially enhanced efficacy for targeted cell ablation. Nature methods, 19(2), 205–215. https://doi.org/10.1038/s41592-021-01364-4
Disciplines
Medicine and Health Sciences
PubMedID
35132245
Department(s)
Department of Obstetrics and Gynecology, Department of Obstetrics and Gynecology Residents, Fellows and Residents
Document Type
Article