NTR 2.0: a rationally engineered prodrug-converting enzyme with substantially enhanced efficacy for targeted cell ablation
Jeff S. Mumm,Mohammed Serhir +1 more
TLDR
In this article , the authors 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.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. read more
Citations
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Synthetic mammalian signaling circuits for robust cell population control
TL;DR: Paradaux as mentioned in this paper is a paradoxical population control circuit, in which the plant hormone auxin stimulates and inhibits net cell growth at different concentrations, and this circuit limited population size over extended timescales of up to 42 days of continuous culture.
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A robust and tunable system for targeted cell ablation in developing embryos.
Z Labbaf,Kleio Petratou,Laura Ermlich,Wilko Backer,Katsiaryna Tarbashevich,Michal Reichman-Fried,Stefan Luschnig,Stefan Schulte-Merker,Erez Raz +8 more
TL;DR: In this article , a cell-ablation system based on bacterial toxin/anti-toxin proteins was developed to enable rapid and cell-autonomous elimination of specific cell types and organs in zebrafish embryos.
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Construction of a Reduction-responsive DNA Microsphere using a Reduction-cleavable Spacer based on a Nitrobenzene Scaffold.
Sayuri L. Higashi,Ayaka Isogami,Jun Takahashi,Aya Shibata,Koichiro M. Hirosawa,Kenichi G. N. Suzuki,Shunsuke Sawada,Shinya Tsukiji,Kazunori Matsuura,Masato Ikeda +9 more
TL;DR: It is demonstrated that the introduction of the RCS in the middle of an oligonucleotide enables the construction of a self-assembled microsphere capable of exhibiting a reduction-responsive disassembly.
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Endoderm-derived islet1-expressing cells differentiate into endothelial cells to function as the vascular HSPC niche in zebrafish.
Hiroyuki Nakajima,Hiroyuki Ishikawa,Takuya Yamamoto,Ayano Chiba,Hajime Fukui,Keisuke Sako,Moe Fukumoto,Kenny Mattonet,Hyouk Bum Kwon,Subhra Prakash Hui,Gergana Dobreva,Kazu Kikuchi,Christian S. M. Helker,Didier Y.R. Stainier,Naoki Mochizuki +14 more
TL;DR: In this paper , the authors identify an unexpected origin for part of the vascular HSPC niche by combining live-imaging-based lineage tracing and single-cell transcriptomics in zebrafish embryos.
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The Role of Transposable Elements in Sexual Development
TL;DR: An overview of the involvement of TEs in processes related to sexual development- from TE activity in the germline to TE accumulation in sex chromosomes is provided and the epigenetic mechanisms that may govern TEs’ role in sexual development are speculated.
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Conditional targeted cell ablation in zebrafish: a new tool for regeneration studies
Silvia Curado,Ryan M. Anderson,Benno Jungblut,Jeff S. Mumm,Eric H. Schroeter,Didier Y.R. Stainier +5 more
TL;DR: The findings show that the NTR/Mtz system is effective for temporally and spatially controlled cell ablation in zebrafish, thereby constituting a most promising genetic tool to analyze tissue interactions as well as the mechanisms underlying regeneration.