The AKAP11 Knockout HEK293T Polyclonal Cells are a heterogeneous population of HEK293T cells carrying a CRISPR/Cas9-mediated disruption of the AKAP11 gene. This polyclonal knockout model circumvents clonal selection artifacts, providing a robust loss-of-function system for interrogating AKAP11-dependent signaling pathways across a diverse genetic background. Unlike knockdown approaches, complete gene disruption ensures thorough interrogation of scaffold functions and downstream effects.
The HEK293T cell line originates from human embryonic kidney epithelium and was immortalized by stable expression of the SV40 large T antigen. This allows episomal amplification of plasmids bearing the SV40 origin, making the line a preferred choice for high-yield protein expression and lentivirus production. HEK293T cells exhibit rapid growth and retain functional cAMP/PKA and Wnt signaling cascades, which are directly relevant to AKAP11 biology.
AKAP11 encodes an A-kinase anchoring protein that scaffolds PKA, PP1, and GSK3?? into multimeric signaling hubs. These complexes integrate upstream cAMP signals, transduced via GPCRs, with Wnt pathway inputs. Anchored PKA phosphorylates downstream effectors such as CREB, while AKAP11-associated GSK3?? regulates ??-catenin stability and glycogen metabolism. Interacting factors including PDE4 and 14-3-3 fine-tune these events. Through coordinated phosphorylation control, AKAP11 modulates gene transcription, cell cycle progression, and metabolic responses.
Disruption of AKAP11 in HEK293T cells abolishes the organized assembly of PKA/PP1/GSK3?? modules, deregulating downstream targets like CREB and ??-catenin. As a polyclonal population, these cells reflect an average knockout phenotype that reduces biases from single clones, making them ideal for bulk biochemical analyses and phenotypic screens. This model is particularly suited for studying how scaffold loss alters cAMP/Wnt crosstalk and impacts proliferation in a transformed cellular context.
Applications include dissecting compartmentalized cAMP/PKA signaling, examining Wnt pathway interplay, modeling neuropsychiatric conditions (e.g., bipolar disorder, schizophrenia), probing cancer cell signaling, and performing kinase inhibitor screens. Compatible techniques encompass western blotting for PKA substrate phosphorylation, co-immunoprecipitation of remaining complex members, immunofluorescence localization, CREB/??-catenin reporter assays, flow cytometry for cell cycle, drug sensitivity testing, and RT-qPCR for target genes. For ordering and support, contact Ascent Research.
