This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population targeting the CD14 gene in the human 786-O renal cell adenocarcinoma line. The CD14 gene disruption was achieved using CRISPR/Cas9 genome editing to generate a heterogeneous pool of cells with loss-of-function modifications, providing a versatile loss-of-function model without single-cell cloning. This polyclonal knockout format preserves biological variability while eliminating CD14-mediated signaling, enabling robust population-level assays in an isogenic background.
The host cell line, 786-O, was originally derived from a primary clear cell renal cell carcinoma (ccRCC) and harbors a well-characterized VHL mutation. This genetic background renders the cells representative of VHL-mutant ccRCC, a major subtype of kidney cancer characterized by constitutive HIF pathway activation. The 786-O line is widely adopted as a model for studying hypoxia-independent oncogenic signaling, tumor metabolism, and the tumor microenvironment in renal cell carcinoma.
CD14 functions as a high-affinity co-receptor for bacterial lipopolysaccharide (LPS), acting upstream of the TLR4/MD-2 complex to initiate innate immune responses. Upon LPS binding, CD14 transfers the ligand to the TLR4/MD-2 complex, engaging the adaptor MyD88 and kinases IRAK4 and TRAF6, which culminates in activation of the NF-??B and MAPK pathways. This signaling cascade promotes transcription of pro-inflammatory cytokines such as TNF, IL6, and IL1B. Upstream regulators of CD14 expression include LPS, lipoproteins, IFN-??, TNF-??, and transcription factors Sp1, AP-1, and NF-??B. CD14 also interacts with LBP, CD11b, and Src family kinases, situating it at a critical juncture in Toll-like receptor and NF-??B signaling.
In the context of 786-O cells, CD14 knockout abrogates LPS-induced inflammatory signaling, providing a clean background to dissect the contribution of CD14-dependent pathways to renal cancer biology. Given the emerging link between chronic inflammation and ccRCC progression, this model allows investigation of how CD14-mediated innate immune recognition influences tumor cell behavior, including proliferation, migration, and cytokine secretion. The VHL-mutant background further enables examination of crosstalk between hypoxia-driven pathways and inflammatory signals, which is relevant to the tumor microenvironment in clear cell kidney cancer.
This polyclonal knockout cell pool is suitable for a wide range of functional studies, including evaluation of LPS-driven tumor progression, screening of anti-inflammatory compounds, and examining the role of CD14 in immune evasion within the tumor microenvironment. Recommended assays include flow cytometry to confirm CD14 loss, ELISA for secreted TNF and IL-6, NF-??B luciferase reporter assays, Western blotting for phosphorylated p65 (ReIA), and RT-qPCR for cytokine transcripts. Migration and invasion assays can be employed to assess phenotypic consequences of disrupted CD14 signaling. For further technical specifications or to discuss customized applications, please contact Ascent Research.
