State Key Laboratory of Medicinal Chemical Biology and College of Pharmacy, Nankai University, Tianjin, 300350, China.
Department of Obstetrics, Tianjin First Central Hospital, Nankai University, Tianjin, 300192, China.
Cell Death Dis. 2022 Mar 30;13(3):284. doi: 10.1038/s41419-022-04738-4.
Reactive oxygen species (ROS) are extensively assessed in physiological and pathological studies; however, the genes and mechanisms involved in antioxidant reactions are elusive. To address this knowledge gap, we used a forward genetic approach with mouse haploid embryonic stem cells (haESCs) to generate high-throughput mutant libraries, from which numerous oxidative stress-targeting genes were screened out. We performed proof-of-concept experiments to validate the potential inserted genes. Slc25a43 (one of the candidates) knockout (KO) ESCs presented reduced damage caused by ROS and higher cell viability when exposed to HO. Subsequently, ROS production and mitochondrial function analysis also confirmed that Slc25a43 was a main target gene of oxidative toxicity. In addition, we identified that KO of Slc25a43 activated mitochondria-related genes including Nlrx1 to protect ESCs from oxidative damage. Overall, our findings facilitated revealing target genes of oxidative stress and shed lights on the mechanism underlying oxidative death.
活性氧 (ROS) 在生理和病理研究中被广泛评估；然而，参与抗氧化反应的基因和机制尚不清楚。为了解决这一知识空白，我们使用具有小鼠单倍体胚胎干细胞 (haESC) 的正向遗传方法生成了高通量突变文库，从中筛选出许多针对氧化应激的靶基因。我们进行了验证潜在插入基因的概念验证实验。当暴露于 HO 时，Slc25a43（候选基因之一）敲除 (KO) ESC 呈现出减少的 ROS 引起的损伤和更高的细胞活力。随后，ROS 产生和线粒体功能分析也证实 Slc25a43 是氧化毒性的主要靶基因。此外，我们发现 Slc25a43 的 KO 激活了包括 Nlrx1 在内的与线粒体相关的基因，以保护 ESC 免受氧化损伤。总的来说，我们的研究结果揭示了氧化应激的靶基因，并阐明了氧化死亡的机制。
