NTR 2.0: a rationally engineered prodrug-converting enzyme with substantially enhanced efficacy for targeted cell ablation
| Autor: | Abigail V. Sharrock, Timothy S. Mulligan, Kelsi R. Hall, Elsie M. Williams, David T. White, Liyun Zhang, Kevin Emmerich, Frazer Matthews, Saumya Nimmagadda, Selena Washington, Katherine D. 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, Jeff S. Mumm |
|---|---|
| Rok vydání: | 2022 |
| Předmět: |
Technology
Green Fluorescent Proteins Genetically Modified CHO Cells Protein Engineering Regenerative Medicine Biochemistry Medical and Health Sciences Retina Animals Genetically Modified Cricetulus Metronidazole Animals Humans Prodrugs Molecular Biology Zebrafish Vibrio 5.2 Cellular and gene therapies Cell Biology Nitroreductases Biological Sciences Stem Cell Research Recombinant Proteins HEK293 Cells Stem Cell Research - Nonembryonic - Non-Human Development of treatments and therapeutic interventions Biotechnology Developmental Biology |
| Zdroj: | Nature methods, vol 19, iss 2 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|