小柯机器人

美国约翰霍普金斯大学Jeff S. Mumm和新西兰惠灵顿维多利亚大学David F. Ackerley共同合作取得重要工作进展。他们开发出了能够显著增强靶向细胞消融功效的NTR 2.0系统。相关论文于2022年2月7日在线发表于《自然—方法学》杂志上。

在这里，研究人员使用合理的工程改造和跨物种筛选开发出了NTR变体，NTR 2.0，它在 MTZ 介导的细胞特异性消融功效方面比NTR提高了约100倍，从而消除了对毒性前药治疗方案的需要。因此，NTR 2.0 能够实现持续的细胞丢失模式和先前抗性细胞类型的消融。这些特性允许增强对细胞功能的探索，对离散干细胞生态位的再生能力提出更大的挑战，以及对慢性退行性疾病进行新的建模。因此，我们创建了一系列双向转基因报告系统资源，以促进 NTR 2.0向研究界的传播。

据介绍，细菌硝基还原酶 (NTR) 的转基因表达使真核细胞对甲硝唑 (MTZ) 等前药敏感，从而实现选择性细胞消融范式，扩大了脊椎动物细胞功能和再生的研究。然而，第一代 NTR 需要混杂有毒的前药治疗才能实现有效的细胞消融，并且某些细胞类型已被证明具有抗药性。

附：英文原文

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

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

Issue&Volume: 2022-02-07

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. An engineered bacterial nitroreductase, NTR 2.0, improves chemically induced cell ablation, facilitating novel sustained ablation paradigms for testing the effects of chronic inflammation on regeneration, and modeling degenerative disease.

DOI: 10.1038/s41592-021-01364-4

Source: https://www.nature.com/articles/s41592-021-01364-4

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex