The ISOC2 Knockout NCI-H1975 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population for investigating ISOC2 function in human lung adenocarcinoma. Transient co-expression of Cas9 endonuclease and locus-specific guide RNAs generates a heterogeneous pool of ISOC2-disrupted alleles, resulting in a loss-of-function model without single-cell cloning. This approach maintains the genetic diversity of the parental NCI-H1975 line while achieving efficient target gene ablation, enabling robust population-level phenotyping.
The parental NCI-H1975 cell line is a human lung adenocarcinoma epithelial model that endogenously expresses EGFR with L858R and T790M point mutations. These dual mutations are key drivers of oncogenic signaling and are associated with acquired resistance to first-generation EGFR tyrosine kinase inhibitors. NCI-H1975 is widely utilized in NSCLC research for studying EGFR-dependent proliferation, survival pathways, and resistance mechanisms. Introducing an ISOC2 knockout into this background creates an isogenic platform to probe the interplay between a poorly characterized metabolic enzyme and mutant EGFR-driven signaling.
ISOC2 encodes a protein bearing an isochorismatase-like domain, which is typical of hydrolases involved in secondary metabolite processing in bacteria; however, its mammalian function remains largely enigmatic. It is predicted to act as a hydrolase and has been tentatively linked to mitochondrial metabolism, but validated substrates, interacting partners, upstream activators, and downstream effectors have not been identified. The ISOC2 signaling network is therefore undefined, positioning this knockout model as a vital tool for deorphanizing the enzyme and elucidating its molecular interactions.
The combination of ISOC2 disruption with the NCI-H1975 EGFR-mutant background offers a unique opportunity to examine metabolic dependencies in lung adenocarcinoma. Constitutive EGFR signaling often drives metabolic reprogramming, including enhanced mitochondrial respiration and altered redox homeostasis. By abrogating ISOC2 function, researchers can assess its contribution to mitochondrial bioenergetics, apoptotic threshold, and metabolic adaptation under oncogenic stress. This model may also reveal synthetic vulnerabilities with EGFR inhibitors, potentially informing strategies to overcome T790M-mediated drug resistance.
This polyclonal knockout population is well-suited for a range of functional assays, including western blotting and RT-qPCR to verify ISOC2 depletion, MTT or CellTiter-Glo viability assays, Annexin V-based apoptosis analysis by flow cytometry, Seahorse mitochondrial stress testing, colony formation assays, and wound healing or Transwell migration/invasion experiments. Drug sensitivity profiling with EGFR inhibitors and chemotherapy agents can assess ISOC2??s role in treatment response. For detailed protocols or commercial inquiries, please contact Ascent Research.
