Development and Application of the CRISPR-dcas13d-eIF4G Translational Regulatory System to Inhibit Ferroptosis in Calcium Oxalate Crystal-Induced Kidney Injury.
Ziqi HeChao SongSheng LiCaitao DongWenbiao LiaoYunhe XiongSixing YangYuchen LiuPublished in: Advanced science (Weinheim, Baden-Wurttemberg, Germany) (2024)
The CRISPR-Cas system, initially for DNA-level gene editing and transcription regulation, has expanded to RNA targeting with the Cas13d family, notably the RfxCas13d. This advancement allows for mRNA targeting with high specificity, particularly after catalytic inactivation, broadening the exploration of translation regulation. This study introduces a CRISPR-dCas13d-eIF4G fusion module, combining dCas13d with the eIF4G translation regulatory element, enhancing target mRNA translation levels. This module, using specially designed sgRNAs, selectively boosts protein translation in targeted tissue cells without altering transcription, leading to notable protein expression upregulation. This system is applied to a kidney stone disease model, focusing on ferroptosis-linked GPX4 gene regulation. By targeting GPX4 with sgRNAs, its protein expression is upregulated in human renal cells and mouse kidney tissue, countering ferroptosis and resisting calcium oxalate-induced cell damage, hence mitigating stone formation. This study evidences the CRISPR-dCas13d-eIF4G system's efficacy in eukaryotic cells, presenting a novel protein translation research approach and potential kidney stone disease treatment advancements.
Keyphrases
- crispr cas
- genome editing
- induced apoptosis
- cell cycle arrest
- cell death
- transcription factor
- genome wide
- endothelial cells
- high glucose
- cancer therapy
- binding protein
- cell proliferation
- signaling pathway
- dna methylation
- gene expression
- single cell
- stem cells
- risk assessment
- poor prognosis
- cell therapy
- case report
- replacement therapy
- combination therapy