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Phenotypic CRISPR screens identify NLRX1 as an essential activator of the human mitochondrial permeability transition - PubMed

6 hours ago
  • #Mitochondria
  • #NLRX1
  • #CRISPR
  • Phenotypic CRISPR screens identified NLRX1 as an essential activator of the human mitochondrial permeability transition (mPT).
  • Two phenotypic assays for mitochondrial calcium overload were designed and applied to FACS-based CRISPR screening in human cells, evaluating 19,113 genes.
  • The first screen studied mitochondrial membrane potential (MMP) collapse in response to calcium overload, identifying MCU and EMRE as top-ranked genes.
  • The second screen measured inner mitochondrial membrane permeability, identifying NF2, REST, BPTF, and NLRX1 as genes affecting mPT.
  • NLRX1 knockout increased mitochondrial calcium retention capacity (CRC), abolished calcium release, and was cyclosporin A (CsA)-insensitive, meeting the definition of an essential mPT activator.
  • Integral membrane proteins, including previously proposed mPT candidates, were not essential activators.