These BRAF knockout 769-P polyclonal cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human clear cell renal cell carcinoma line 769-P, engineered to disrupt the gene encoding the serine/threonine kinase BRAF. This polyclonal format provides a heterogeneous pool of clones with diverse loss-of-function mutations, facilitating comprehensive studies of BRAF-deficient signaling in renal carcinoma models.
The parental 769-P line is a VHL wild-type clear cell renal cell carcinoma (ccRCC) model extensively used to investigate kidney cancer biology independently of VHL inactivation. These cells retain key signaling pathways relevant to ccRCC pathogenesis, making them an ideal background for interrogating BRAF function. The 769-P line displays dysregulated growth factor signaling and sensitivity to kinase inhibitors, providing a physiologically relevant system for BRAF knockout studies.
BRAF encodes a serine/threonine kinase that functions as a central node in the RAS-RAF-MEK-ERK MAPK cascade. Activated by upstream regulators including RAS GTPases (HRAS, KRAS, NRAS) and receptor tyrosine kinases, BRAF directly phosphorylates MEK1/2, which then phosphorylate ERK1/2. Activated ERKs regulate transcription factors such as ELK1, c-MYC, and Cyclin D1 to drive proliferation and survival. BRAF interacts with scaffold proteins like KSR, chaperones such as Hsp90, and other RAF family members (ARAF, CRAF). Disruption of BRAF expression abolishes downstream signaling, enabling dissection of pathway dependencies and compensatory mechanisms.
In VHL wild-type ccRCC, these polyclonal knockout cells facilitate investigation of MAPK pathway addiction and resistance mechanisms independent of the VHL-HIF axis. Researchers can examine how loss of BRAF influences ERK phosphorylation, cell cycle progression, and apoptosis specifically within a renal carcinoma background. This model also permits study of compensatory activation of CRAF or PI3K-AKT signaling, relevant to acquired inhibitor resistance.
These BRAF knockout polyclonal cells are suited for signaling studies using Western blotting for phospho-MEK/ERK, drug sensitivity assays with MAPK inhibitors (vemurafenib, dabrafenib, trametinib), and functional assays like proliferation (MTT) and flow cytometry for cell cycle/apoptosis. Transcriptome profiling via RNA-seq and clonogenic survival assays further define pathway rewiring and resistance mechanisms. The polyclonal nature captures a broad spectrum of mutations, providing a comprehensive view of functional outcomes. For additional information or custom projects, please contact Ascent Research.
