The AZGP1 Knockout HEK293T Polyclonal Cells product is a ready-to-use CRISPR/Cas9-edited polyclonal knockout cell population engineered to disrupt the AZGP1 gene. This loss-of-function model eliminates zinc-alpha-2-glycoprotein (ZAG) secretion, providing a defined null background for investigating ZAG-mediated lipolytic signaling. The polyclonal format captures a heterogeneous pool of edited cells, offering a robust and rapid resource for functional studies without requiring clonal isolation.
HEK293T cells are a derivative of the HEK293 human embryonic kidney epithelial cell line, stably expressing the SV40 large T antigen. This modification enhances episomal replication of plasmids containing the SV40 origin of replication, making HEK293T a widely used host for high-level transient protein expression and lentiviral/retroviral production. Their epithelial origin and robust growth characteristics render them amenable to a broad range of genetic manipulations and functional assays, including CRISPR/Cas9-mediated gene knockout.
AZGP1 encodes zinc-alpha-2-glycoprotein (ZAG), a lipid-mobilizing factor that promotes lipolysis via beta-adrenergic receptor signaling, primarily through ADRB3. ZAG secretion is regulated by upstream signals including glucocorticoids, androgens, and beta-adrenergic agonists. In the lipolytic cascade, ZAG engagement with ADRB3 activates adenylyl cyclase, elevating cAMP and activating PKA. PKA phosphorylates hormone-sensitive lipase (HSL) and modulates ATGL activity, resulting in triglyceride hydrolysis into free fatty acids. Knockout of AZGP1 in HEK293T cells disrupts this signaling axis, enabling precise dissection of ZAG??s role in lipid metabolism and its interactions with free fatty acid feedback.
Although HEK293T cells are non-adipocytic, they provide an effective platform for studying ZAG secretion and paracrine signaling. The polyclonal knockout population supports experiments such as ZAG-conditioned media transfer to adipocytes, co-culture with cancer cell lines, and high-throughput screening for modulators of ZAG release or receptor binding. This model is particularly valuable for investigating mechanisms underlying cancer cachexia, where ZAG-driven lipolysis contributes to adipose wasting.
Representative assays include Western blot and ELISA for ZAG, RT-qPCR targeting AZGP1, cAMP accumulation assays, and lipolysis measurements in recipient adipocytes. The knockout cells are suitable for functional rescue, ADRB3 co-immunoprecipitation, and genetic screens. These cells provide a consistent model for metabolic and cancer cachexia research. For pricing and availability, contact Ascent Research.
