Genome-wide screening in the haploid system reveals Slc25a43 as a target gene of oxidative toxicity.
Jinxin ZhangYiding ZhaoYaru TianMengyang GengYan LiuWenhao ZhangLing ShuaiPublished in: Cell death & disease (2022)
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 H 2 O 2 . 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.
Keyphrases
- genome wide
- oxidative stress
- reactive oxygen species
- embryonic stem cells
- dna methylation
- dna damage
- genome wide identification
- copy number
- cell death
- high throughput
- ischemia reperfusion injury
- induced apoptosis
- diabetic rats
- genome wide analysis
- healthcare
- bioinformatics analysis
- transcription factor
- single cell
- risk assessment