Recanalization therapy by intravenous thrombolysis or endovascular therapy is critical for the treatment of cerebral infarction. However, the recanalization treatment will also exacerbate acute brain injury and even severely threatens human life due to the reperfusion injury. So far, the underlying mechanisms for cerebral ischaemia‐reperfusion injury are poorly understood and effective therapeutic interventions are yet to be discovered. Therefore, in the research, we subjected SK‐N‐BE(2) cells to oxygen‐glucose deprivation/reperfusion (OGDR) insult and performed a pooled genome‐wide CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR‐associated protein 9) knockout screen to discover new potential therapeutic targets for cerebral ischaemia‐reperfusion injury. We used Metascape to identify candidate genes which might involve in OGDR resistance. We found that the genes contributed to OGDR resistance were primarily involved in neutrophil degranulation, mitochondrial translation, and regulation of cysteine‐type endopeptidase activity involved in apoptotic process and response to oxidative stress. We then knocked down some of the identified candidate genes individually. We demonstrated that MRPL19, MRPL32, MRPL52 and MRPL51 inhibition increased cell viability and attenuated OGDR‐induced apoptosis. We also demonstrated that OGDR down‐regulated the expression of MRPL19 and MRPL51 protein. Taken together, our data suggest that genome‐scale screening with Cas9 is a reliable tool to analyse the cellular systems that respond to OGDR injury. MRPL19 and MRPL51 contribute to OGDR resistance and are supposed to be promising targets for the treatment of cerebral ischaemia‐reperfusion damage.

中文翻译：

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CRISPR/Cas9 介导的全基因组全基因敲除筛选鉴定了参与氧糖剥夺/再灌注抵抗的线粒体核糖体蛋白。


静脉溶栓或血管内治疗的再通治疗对于脑梗塞的治疗至关重要。然而，再通治疗也会加剧急性脑损伤，甚至因再灌注损伤而严重威胁人类生命。迄今为止，人们对脑缺血再灌注损伤的潜在机制知之甚少，有效的治疗干预措施尚未发现。因此，在研究中，我们对 SK-N-BE(2) 细胞进行了氧葡萄糖剥夺/再灌注 (OGDR) 损伤，并进行了全基因组 CRISPR（成簇规则间隔短回文重复序列）/Cas9（CRISPR 相关蛋白9）敲除筛选，发现脑缺血再灌注损伤新的潜在治疗靶点。我们使用 Metascape 来鉴定可能涉及 OGDR 抗性的候选基因。我们发现导致 OGDR 抵抗的基因主要涉及中性粒细胞脱颗粒、线粒体翻译以及参与细胞凋亡过程和氧化应激反应的半胱氨酸型内肽酶活性的调节。然后我们单独敲除一些已识别的候选基因。我们证明，抑制 MRPL19、MRPL32、MRPL52 和 MRPL51 可增加细胞活力并减弱 OGDR 诱导的细胞凋亡。我们还证明 OGDR 下调 MRPL19 和 MRPL51 蛋白的表达。综上所述，我们的数据表明，使用 Cas9 进行基因组规模筛选是分析响应 OGDR 损伤的细胞系统的可靠工具。 MRPL19 和 MRPL51 有助于 OGDR 抵抗，被认为是治疗脑缺血再灌注损伤的有希望的靶点。