The ANKS1B Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that provides a loss-of-function model for the ANKS1B gene in the widely used HEK293T host cell line. As a heterogeneous pool of edited cells, this product enables functional interrogation of ANKS1B-dependent signaling without the selection of single-cell clones, offering a representative population-level knockout response. This tool is designed for studies of receptor tyrosine kinase signaling, particularly the ephrin/Eph pathway, and is suitable for a broad range of biochemical and cell-based assays.
The HEK293T cell line is a derivative of human embryonic kidney epithelial cells that stably expresses the SV40 large T antigen, facilitating high-copy episomal replication of plasmids containing the SV40 origin of replication and resulting in robust transient protein expression. This host cell line is extensively utilized for protein production, signaling pathway reconstitution, and functional genomics applications. Its adherent epithelial morphology supports standard cell culture, transfection, and imaging protocols, making it compatible with immunofluorescence, migration, and reporter assays that are central to studying scaffold proteins like ANKS1B.
ANKS1B encodes a cytoplasmic scaffold protein that functions downstream of the EphA8 receptor tyrosine kinase. Upon binding of ephrin-A ligands, EphA8 undergoes autophosphorylation, creating docking sites for ANKS1B. Phosphorylated ANKS1B interacts with the adaptor proteins Grb2 and Gab1, nucleating signaling complexes that propagate signals through the MAPK and PI3K-Akt cascades. This leads to activation of ERK1/2 and Akt kinases, which in turn modulate Rho GTPases and cytoskeletal regulators to influence cell adhesion and migration. Additionally, ANKS1B-mediated signals can alter gene expression via transcription factors such as c-Fos, linking extracellular ephrin cues to changes in neuronal connectivity and synaptic function.
In the HEK293T background, the ANKS1B knockout disrupts the canonical EphA8-to-MAPK/PI3K-Akt signaling axis, providing a clean cellular system for dissecting the molecular contributions of this scaffold. The absence of endogenous ANKS1B allows for structure-function studies through transient complementation with wild-type or mutant ANKS1B constructs, enabling mapping of interaction domains required for Grb2 or Gab1 recruitment. The polyclonal nature of the knockout population mimics biological variability and avoids artifacts that may arise from clonal selection, making it suitable for robust phenotypic screening and dose-response experiments.
Researchers can employ this knockout model to investigate Eph-ephrin signaling in neurodevelopmental contexts, assess the role of ANKS1B in synaptic plasticity, and model aspects of neurodevelopmental disorders such as intellectual disability, autism spectrum disorder, and schizophrenia. Typical applications include Western blot analysis of phosphorylated ERK1/2, RT-qPCR profiling of downstream target genes, immunofluorescence detection of EphA8 clustering, and cell migration assays. Furthermore, the cell population can be used for high-throughput screening of small molecules that modulate receptor tyrosine kinase pathways. For additional technical specifications, custom product inquiries, or collaborative opportunities, please contact Ascent Research.
