The BRAF Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the BRAF gene in the HCT 116 human colorectal carcinoma cell line. This product provides a heterogeneous pool of cells with BRAF gene disruption, enabling rapid functional studies of BRAF-dependent phenotypes without clonal isolation. These knockout cells serve as a versatile tool for loss-of-function investigations in a well-defined oncogenic background and are compatible with standard cellular and biochemical assays.
HCT 116 is a widely used colorectal carcinoma epithelial cell line carrying a KRAS G13D mutation. This mutation activates the RAS-RAF-MEK-ERK signaling cascade constitutively, making the line an established model for colorectal cancer research, drug screening, and oncogenic signaling studies. HCT 116 cells exhibit stable in vitro growth and are readily transfectable, facilitating genetic manipulation and downstream analyses.
BRAF encodes a serine/threonine kinase that acts downstream of RAS GTPases to phosphorylate and activate MEK1/2, triggering ERK1/2 phosphorylation and nuclear translocation. Active ERK phosphorylates ELK1, inducing expression of proliferation and survival genes such as c-FOS, c-JUN, cyclin D1, and MYC. BRAF activation is promoted by RAS-GTP binding, dimerization with ARAF or RAF1, and interactions with scaffold proteins KSR1/2, co-chaperones CDC37/HSP90, and 14-3-3 proteins. Upstream signals include receptor tyrosine kinases (EGFR, FGFR), growth factors (EGF, FGF), and RAS family members (KRAS, NRAS, HRAS). This kinase thus occupies a central node in the MAPK/ERK pathway, linking extracellular cues to transcriptional programs governing proliferation, differentiation, and survival.
In HCT 116, oncogenic KRAS drives constitutive BRAF-dependent ERK signaling. Disrupting BRAF in this context can reveal the extent to which mutant KRAS relies on BRAF versus other RAF isoforms, and may uncover adaptive signaling rewiring upon RAF inhibition. The polyclonal knockout population captures cellular heterogeneity, reflecting potential variable responses to pathway inhibitors and mimicking the diversity observed in tumors. These cells are therefore a valuable model for studying resistance mechanisms, identifying bypass pathways, and evaluating combination therapy strategies against the RAS-RAF-MEK-ERK cascade.
Typical research applications include western blot analysis of phospho-ERK, RT-qPCR measurement of downstream targets (c-FOS, cyclin D1), proliferation and colony formation assays, and drug sensitivity profiling with BRAF or MEK inhibitors. Additional assays such as flow cytometry for apoptosis/cycle, phospho-kinase arrays, and signaling network analysis can be performed. These BRAF knockout cells support colorectal cancer biology, targeted therapy resistance, and MAPK pathway signaling studies. For further details, please contact Ascent Research.
