The AKAP11 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HT29 colorectal adenocarcinoma cells, engineered to disrupt the AKAP11 gene. This heterogeneous population provides a loss-of-function model for studying AKAP11??s role in compartmentalized cAMP signaling and colorectal cancer biology. The polyclonal format captures genetic diversity from CRISPR/Cas9-mediated gene disruption, enabling bulk cell studies without clonal selection bias. These cells serve as a robust tool for functional genomics, drug screening, and pathway analysis.
The HT29 host cell line originates from a primary colorectal adenocarcinoma of a 44-year-old female and exhibits epithelial adherent morphology with retained colonocyte characteristics. Under appropriate culture conditions, HT29 cells can undergo enterocytic differentiation, making them a well-established model for intestinal epithelial biology and colorectal cancer research. The cell line expresses key colorectal cancer markers and signaling components, and it is widely employed to study tumorigenesis, metastasis, and drug responses. Its adherent nature facilitates a range of downstream assays, including imaging, biochemical fractionation, and high-content screening.
AKAP11 is a dual-specificity A-kinase anchoring protein that scaffolds PKA regulatory subunits (RII??/??) and PP1 catalytic subunit, thereby compartmentalizing cAMP/PKA signaling. By controlling the phosphorylation of ??-catenin and LATS1, AKAP11 modulates both Wnt and Hippo pathway outputs. AKAP11-bound PP1 dephosphorylates LATS1, promoting YAP/TAZ nuclear translocation and transcriptional activity, while AKAP11 regulation of ??-catenin stability influences TCF/LEF-dependent gene expression. Additional interactions with GSK3?? and BAD connect AKAP11 to cell cycle progression and apoptosis. Consequently, AKAP11 represents a convergence point for cAMP/PKA, Hippo, and Wnt/??-catenin signaling in controlling cellular fate.
In colorectal cancer, AKAP11 knockout in HT29 cells provides a model to dissect compartmentalized PKA signaling in oncogenic proliferation and survival. HT29 cells harbor APC and TP53 mutations, mimicking colorectal cancer genetics, and AKAP11 deletion enables study of PKA-anchoring dependency in a tumor-relevant background. Loss of AKAP11 is expected to perturb YAP/TAZ and ??-catenin/TCF transcriptional balance, potentially affecting cyclin D1 and apoptotic thresholds. This model is relevant for investigating tumor maintenance and synthetic vulnerabilities linked to PKA anchoring in colorectal cancer.
These polyclonal knockout cells enable AKAP11-dependent Hippo and Wnt pathway analysis via ??-catenin/TCF reporter and phospho-PKA substrate assays, cell viability and apoptosis measurements, colony formation, migration/invasion studies, and co-immunoprecipitation for protein complex mapping. They are also suitable for drug screening to identify compounds with differential effects on AKAP11-proficient versus -deficient CRC cells. For further technical inquiries or to discuss specific experimental needs, please contact Ascent Research.
