The CBL Knockout MES-OV Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from MES-OV mouse embryonic stem cells, in which the Cbl gene has been disrupted to ablate expression of the E3 ubiquitin-protein ligase CBL. This polyclonal product bypasses clonal selection, providing a heterogeneous loss-of-function model that captures diverse editing events and facilitates population-level studies of signaling dynamics and cellular phenotypes.
MES-OV cells originate from the 129/SvJ mouse strain and retain pluripotent embryonic stem cell characteristics, including self-renewal capacity and the ability to differentiate into ectoderm, mesoderm, and endoderm derivatives. They are extensively utilized in developmental biology for directed differentiation protocols, gene targeting experiments, and genome-wide functional screens, and their germline-competent background enables the generation of transgenic mouse models and exploration of early embryonic signaling mechanisms.
CBL is a RING-type E3 ubiquitin ligase that, upon recruitment to activated receptor tyrosine kinases (RTKs) via adaptor proteins Grb2 and CrkL, mediates K48-linked ubiquitination of targets such as EGFR, PDGFR, CSF-1R, and MET, marking them for proteasomal degradation. It also interacts with CIN85, Sprouty proteins, and the p85 regulatory subunit of PI3K to fine-tune signal termination. Through this activity, CBL negatively regulates the Grb2?CSOS?CRas?CRaf?CMEK?CERK and PI3K?CAKT?CmTOR signaling axes. Consequently, Cbl knockout leads to sustained phosphorylation of downstream effectors including AKT and ERK, amplifying mitogenic and survival signals.
In the pluripotent MES-OV background, loss of CBL function heightens RTK signaling, perturbing the balance between self-renewal and lineage commitment and potentially conferring oncogenic properties akin to those seen in CBL-mutant myeloid malignancies such as chronic myelomonocytic leukemia and juvenile myelomonocytic leukemia. This model also enables dissection of how ubiquitin-dependent receptor downregulation intersects with core pluripotency circuitry and with cell cycle regulators like cyclin D1 and anti-apoptotic Bcl-2 members.
This cell population supports a wide array of research applications: Western blotting for CBL protein and phospho-signaling intermediates (p-EGFR, p-AKT, p-ERK); flow cytometry to assess RTK surface expression; RNA-seq for global transcriptomic analysis; co-immunoprecipitation to validate CBL interactions with Grb2, CrkL, or p85; in vitro ubiquitination assays; proliferation and apoptosis assays; and drug sensitivity testing against MAPK or PI3K pathway inhibitors. For technical specification documents or experimental consultation, please contact Ascent Research.
