This product consists of a polyclonal population of HEK293T cells with CRISPR/Cas9-mediated gene disruption of ANKRD17, generating a heterogeneous loss-of-function model for studying the scaffold protein??s role in tumor suppression and signal transduction. The polyclonal format provides a diverse cellular pool, representing a range of editing outcomes that collectively ablate ANKRD17 function without requiring single-cell cloning, enabling robust analysis in a widely used expression host.
HEK293T cells are derived from human embryonic kidney epithelial cells and stably express the SV40 large T antigen, which promotes episomal replication of plasmids containing the SV40 origin of replication. This immortalized line exhibits adherent growth, high transfectability, and sustained recombinant protein production, making it a foundational model for biochemical reconstitution, transient overexpression, and reporter-based signaling assays across academia and industry.
ANKRD17 encodes a multi-domain scaffold protein characterized by ankyrin repeats that physically bridges key tumor-suppressive pathways. It interacts with TP53 and enhances its transactivation of cell cycle arrest and apoptosis genes such as CDKN1A (p21) and BAX. In the Hippo pathway, ANKRD17 associates with YAP1, LATS1, and 14-3-3 proteins to modulate YAP1 nuclear translocation and TEAD-mediated transcription. Additionally, ANKRD17 contributes to Wnt signaling through interactions with ??-catenin, linking it to TCF/LEF-dependent gene expression. Its activity is responsive to upstream DNA damage signals and LATS1/2-mediated phosphorylation, positioning ANKRD17 at a convergent node for p53, Hippo, and Wnt cascades.
In the HEK293T context, disruption of ANKRD17 provides a versatile platform for dissecting the interplay between these pathways in an epithelial cell background. Although SV40 large T antigen partially abrogates p53 activity, the remaining p53-dependent functions and Hippo regulatory circuits remain measurable using sensitive transcriptional reporters and subcellular localization assays. Polyclonal knockout cells thus facilitate examination of ANKRD17-dependent changes in YAP1 distribution, p53 target expression, and ??-catenin-driven transcription without clonal bias, supporting mechanistic studies into how ANKRD17 coordinates cellular responses to stress and proliferative cues.
Typical applications encompass cancer biology, apoptosis research, Hippo/p53 pathway analysis, and drug screening for modulators of tumor suppression. Experimentally, these cells are suited for Western blotting of p53 targets and phosphorylated YAP, immunofluorescence tracking of YAP1 localization, dual-luciferase reporter assays for p53 or TEAD activity, Annexin V/PI apoptosis assays, co-immunoprecipitation of ANKRD17 complex partners, and RT-qPCR of downstream effectors. For additional technical details, researchers are encouraged to contact Ascent Research.
