铁死亡是一种伴随着大量铁离子积聚和脂质过氧化的细胞调控死亡过程。光激活的核苷酸增强交联和免疫沉淀（PAR-CLIP）是一种用于在核苷酸水平上高分辨率识别RNA结合蛋白（RBPs）与靶向RNA结合位点的方法。通过将光敏核苷酸类似物插入活体细胞中的新RNA转录本中，可以在逆转录过程中产生特征性突变，并用于准确定位RNA和RBPs的交联位置。利用PAR-CLIP检测RNA与RBPs之间的相互作用，确定精确的交联位点，或通过已知蛋白质搜索铁死亡通路基因上游或下游的RNA，可以帮助阐明和验证铁死亡各个信号通路的发生和调控机制。此外，它可能揭示了铁死亡检测的新靶点，提高了铁死亡相关疾病（如癌症和神经退行性疾病）的治疗效率。在这里，我们介绍了一种特定的PAR-CLIP方案来监测铁死亡过程。© 2023. 作者独家许可给Springer Science+Business Media, LLC，属于Springer Nature部分。

Ferroptosis is a regulatory cell death process that is accompanied by large amounts of iron ion accumulation and lipid peroxidation. Photoactivated ribonucleoside-enhanced crosslinking and immunoprecipitation (PAR-CLIP) is a method used to identify the binding sites of RNA-binding proteins (RBPs) on target RNAs with high resolution at the nucleotide level. By inserting photosensitive ribonucleoside analogs into new RNA transcripts of living cells, characteristic mutations can be generated during reverse transcription and be used to accurately locate the crosslinking position of RNAs and RBPs. The use of PAR-CLIP to detect interactions and determine precise crosslinking sites between RNAs and RBPs, or to search for RNAs upstream or downstream of ferroptosis pathways genes through known proteins, can help to clarify and verify the occurrence and regulation mechanisms of the various signaling pathways of ferroptosis. Furthermore, it may reveal new targets for ferroptosis detection and improve the treatment efficiency of ferroptosis-related diseases such as cancer and neurodegenerative diseases. Here, we introduce a specific PAR-CLIP protocol for monitoring the ferroptosis process.© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.