The CD14 Knockout DLD-1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated from the DLD-1 human colorectal adenocarcinoma cell line through targeted disruption of the CD14 gene. This loss-of-function model provides a stable and heterogeneous gene-edited pool, enabling robust investigation of CD14-dependent signaling pathways in an epithelial cancer background. The polyclonal format captures a diverse spectrum of knockout events within a single population, offering a versatile tool for studying CD14-mediated processes without clonal selection artifacts. Researchers can employ this product to examine innate immune receptor function, pattern recognition mechanisms, and downstream inflammatory cascades in a genetically defined colorectal cancer context.
The parental DLD-1 cell line is a well-established human colorectal adenocarcinoma model characterized by endogenous mutations in the adenomatous polyposis coli (APC) and tumor protein p53 (TP53) genes. These oncogenic alterations drive constitutive Wnt pathway activation and impair genomic surveillance, mirroring key features of sporadic colorectal carcinogenesis. DLD-1 cells exhibit an epithelial morphology and retain the capacity for growth factor-dependent proliferation, making them suitable for signal transduction studies. The co-occurrence of APC and TP53 mutations creates a tumor microenvironment-relevant background in which innate immune signaling can be systematically dissected, particularly in the context of microbial sensing and inflammatory responses.
CD14 functions as a glycosylphosphatidylinositol (GPI)-anchored pattern recognition receptor and serves as a critical co-receptor for bacterial lipopolysaccharide (LPS) and lipoteichoic acid. Upon LPS binding, CD14 delivers the ligand to the toll-like receptor 4 (TLR4)/MD-2 complex, triggering myeloid differentiation primary response 88 (MyD88)-dependent signaling that culminates in activation of the transcription factor nuclear factor-??B (NF-??B) and mitogen-activated protein kinases (MAPKs) including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38. This cascade promotes expression of pro-inflammatory cytokines such as tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-1?? (IL-1??). CD14 activity is modulated by upstream regulators including interferon-?? (IFN-??) and the LPS-binding protein (LBP), and CD14 physically interacts with the TLR4/MD-2 complex to facilitate signal initiation. Through these interactions, CD14 acts as a sentinel receptor connecting microbial recognition to intracellular inflammatory programs.
In the DLD-1 colorectal cancer milieu, CD14-mediated innate immune signaling intersects with oncogenic pathways driven by APC and TP53 mutations. The inflammatory response elicited by LPS or other microbial products can influence tumor cell proliferation, survival, and interaction with the immune microenvironment. Ablating CD14 expression in this genetic background allows dissection of how pattern recognition receptor signaling contributes to cancer-associated inflammation and potential tumor-promoting networks. The knockout model is particularly valuable for investigating the role of bacterial sensing in colorectal cancer progression, given the gut??s constant exposure to microbial ligands. By eliminating CD14-dependent signal transduction, researchers can differentiate between direct tumor cell-intrinsic pathways and microenvironmental contributions to inflammatory cytokine production.
This polyclonal CD14 knockout cell population is optimized for a broad range of experimental applications, including mechanistic studies of LPS-induced NF-??B and MAPK signaling, cytokine secretion profiling, and microbial-host interaction analyses. Representative techniques include western blotting for phosphorylated NF-??B p65, enzyme-linked immunosorbent assays (ELISAs) for TNF and IL-6 secretion, NF-??B luciferase reporter assays, flow cytometric analysis of CD14 surface expression to confirm gene disruption, and quantitative reverse transcription PCR (RT-qPCR) for pro-inflammatory cytokine transcripts. The model supports research on sepsis-like inflammatory responses, innate immune checkpoint mechanisms in cancer, and pharmacological modulation of TLR4-dependent pathways. For additional technical details or customer support inquiries, please contact Ascent Research.
