The CCM2 Knockout 786-O Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the human 786-O cell line. This heterogeneous pool of cells carries targeted disruptions in the CCM2 gene, providing a loss-of-function model for studying CCM2-dependent signaling and cellular processes. As a polyclonal population, the product captures diverse allelic knockout events across the pool, making it suitable for population-level assays where monoclonal isolation is not required.
The parental 786-O cell line is a widely used model of clear cell renal cell carcinoma (ccRCC), originally established from a primary tumor of a 58-year-old Caucasian male. These adherent epithelial cells retain tumorigenic properties, including anchorage-independent growth, and express characteristic ccRCC markers such as mutant VHL. Their epithelial origin and junctional competency make them particularly relevant for investigating CCM2-mediated junction integrity in a cancer context.
CCM2 encodes a scaffold protein that forms the core CCM complex with KRIT1 (CCM1) and PDCD10 (CCM3). This complex negatively regulates the MEKK3-MEK5-ERK5 MAPK module and the RhoA/ROCK pathway. Mechanistically, CCM2 bridges KRIT1 and PDCD10 to suppress MEKK3 activation, thereby limiting ERK5 phosphorylation and downstream transcription of KLF2 and KLF4. Concurrently, the complex attenuates RhoA/ROCK/LIMK/cofilin signaling to maintain cortical actin dynamics and cell-cell junction stability. CCM2 also interacts with ICAP-1 and integrin ??1, linking integrin-mediated adhesion to junctional control. Loss of CCM2 disrupts this balance, leading to hyperactive ERK5 and increased RhoA-driven contractility, which compromises endothelial and epithelial barrier function.
In the 786-O epithelial context, CCM2 knockout provides a unique platform to dissect the interplay between the CCM complex and renal cancer biology. While CCM2 mutations are clinically associated with cerebral cavernous malformations, its role in epithelial junction maintenance and MAPK/Rho crosstalk has significant implications for tumor cell behavior, including migration, invasion, and metastasis. The polyclonal knockout pool enables researchers to assess heterogeneous CCM2 loss-of-function effects in a ccRCC background, shedding light on how junctional dysregulation contributes to malignant phenotypes.
Typical applications include western blotting and RT-qPCR to confirm loss of CCM2 and downstream target changes (e.g., KLF2, KLF4, RhoA); immunofluorescence to visualize junction protein mislocalization; cell migration and invasion assays; RhoA activity and ERK5 phosphorylation analyses; and transcriptomic profiling via RNA-seq. The cells also serve as a tool for drug screening to identify inhibitors of aberrant MEKK3-ERK5 or Rho/ROCK signaling, and for defining novel CCM2 interactors. For technical inquiries or ordering, contact Ascent Research.
