LSS Knockout Raji Polyclonal Cells are a genetically engineered human B lymphocyte population featuring CRISPR/Cas9-mediated disruption of the LSS gene, which encodes lanosterol synthase. This polyclonal knockout model provides a heterogeneous loss-of-function system ideally suited for studying cholesterol metabolism and sterol homeostasis without the selection bias of single-cell clones. The targeted gene disruption ablates functional protein expression across the population, enabling robust analysis of LSS-dependent cellular processes in a physiologically relevant immune cell background. Researchers can employ these cells to investigate the immediate metabolic consequences of lanosterol synthase deficiency and downstream compensatory mechanisms within an intact mammalian system.
The Raji host cell line originates from a Burkitt lymphoma patient and is extensively characterized as an Epstein-Barr virus (EBV)-positive, suspension lymphoblastoid line. As a B lymphocyte model, Raji cells inherently perform immune functions such as antibody production and antigen presentation, rendering them exceptionally useful for immuno-oncology and infectious disease research. Their rapid proliferation in culture and well-defined signaling networks make them a workhorse for studying cancer biology, drug responses, and lipid metabolism interventions. The suspension growth format facilitates homogeneous compound exposure and scalability for high-throughput screening.
LSS catalyzes the cyclization of (S)-2,3-oxidosqualene to lanosterol, a critical step in sterol biosynthesis that branches toward cholesterol, steroid hormones, bile acids, and vitamin D production. Expression of LSS is tightly controlled by sterol-sensing machinery: the transcription factors SREBF1 and SREBF2, in concert with the SREBP cleavage-activating protein (SCAP) and INSIG1, upregulate its expression under low-cholesterol conditions. Lanosterol synthetase acts downstream of squalene epoxidase (SQLE) and cooperates with CYP51A1, MSMO1, and HSD17B7 in the post-lanosterol pathway. Disrupted LSS activity leads to accumulation of oxidosqualene precursors and depletion of lanosterol, driving compensatory SREBP activation and altering the expression of enzymes like HMGCR, MVK, PMVK, and DHCR7, thereby rerouting metabolic flux within the isoprenoid and sterol networks.
In the Raji lymphoma context, LSS knockout profoundly perturbs lipid raft assembly, membrane biogenesis, and protein prenylation, each of which is essential for B cell receptor signaling, proliferation, and survival. The resulting cholesterol auxotrophy can expose synthetic lethal vulnerabilities relevant to congenital cataracts, alopecia subtypes, hypercholesterolemia, and metabolic syndrome. This model also empowers exploration of the crosstalk between cholesterol metabolism and oncogenic pathways, as lymphoma cells often exhibit elevated sterol demand. The polyclonal nature of the knockout captures cell-to-cell variability in compensatory responses, mirroring the heterogeneity observed in tumor populations and patient-derived samples.
These cells enable a broad spectrum of applications: validating LSS as a therapeutic target in metabolic disorders and cancer, high-throughput inhibitor screening, and mechanistic dissection of sterol-dependent immune functions. Researchers can confirm target disruption via RT-qPCR and Western blotting, quantify cholesterol levels using the Amplex Red assay, assess proliferation through MTT or BrdU incorporation, and evaluate apoptosis by flow cytometry with Annexin V staining. Lipid droplet accumulation may be visualized with Nile red, and downstream signaling events can be probed via phospho-specific immunoblotting. For custom experimental design or to procure this product, please contact Ascent Research.
