The AHNAK2 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for the functional investigation of AHNAK2, a giant scaffold protein implicated in calcium signaling and actin cytoskeleton dynamics. This genetically disrupted pool of HEK293T cells provides a loss-of-function model that circumvents clonal selection artifacts while maintaining robust, reproducible knockout across the population. The targeted disruption of the AHNAK2 gene enables precise dissection of its roles without the need for single-cell cloning, making this product suitable for high-throughput screening and comparative signaling studies.
The parental HEK293T cell line is a widely utilized human embryonic kidney epithelial derivative that stably expresses the SV40 large T antigen, facilitating high-level episomal replication of plasmids carrying the SV40 origin. Renowned for its exceptional transfectability and protein production capacity, HEK293T serves as a premier host for recombinant protein expression, lentivirus packaging, and biochemical analyses. This cell line??s robust growth characteristics and well-characterized signaling milieu offer a consistent and reproducible platform for interrogating the cellular functions of AHNAK2 in a simplified but biologically relevant context.
AHNAK2 is a large scaffold protein that integrates calcium signals to regulate actin cytoskeleton dynamics, controlling cell adhesion and migration. It directly binds calcium-sensitive proteins S100A10 and Annexin A2, as well as actin and calmodulin, forming a signaling complex at the membrane. Upstream, AHNAK2 is activated by calcium influx and is phosphorylated by PKC and ERK1/2 in response to growth factors. Downstream, it modulates actin filament assembly and adhesion complex formation, bridging calcium signaling to cellular mechanics. This positions AHNAK2 as a key mediator of processes like cell spreading, motility, and mechanotransduction.
In the HEK293T background, AHNAK2 knockout decouples calcium-dependent cytoskeletal regulation from confounding factors, enabling focused study. These cells retain core calcium signaling machinery, including calmodulin and ERK, allowing dissection of AHNAK2-specific effects on actin dynamics and adhesion. The polyclonal knockout population avoids clonal adaptation artifacts, offering an authentic loss-of-function model. Moreover, HEK293T??s high transfection efficiency permits rescue experiments with AHNAK2 mutants for structure-function analyses.
Applications include cancer cell migration and metastasis studies using transwell assays and immunofluorescence for focal adhesions. Drug target validation can be performed by monitoring phospho-ERK via Western blot or calcium imaging after pathway perturbation. Protein interaction analyses via co-immunoprecipitation of partners like S100A10 and Annexin A2 are facilitated. The knockout population also serves as a negative control in lentivirus production or for investigating Wnt?Ccalcium crosstalk. For further information, contact Ascent Research.
