AKAP13 Knockout HT29 Polyclonal Cells are a population of CRISPR/Cas9-edited human colorectal adenocarcinoma cells carrying targeted disruption of the AKAP13 gene. This polyclonal knockout pool, derived from the HT29 cell line, provides a genetically heterogeneous model to investigate the consequences of AKAP13 loss of function in intestinal epithelial biology and colorectal cancer. The use of CRISPR/Cas9-mediated gene disruption enables efficient abrogation of AKAP13 protein expression, facilitating studies of downstream signaling defects without the confounding influences of clonal selection.
The HT29 host cell line is a well-characterized human epithelial colorectal adenocarcinoma line established from a primary tumor. HT29 cells exhibit epithelial morphology and, under appropriate culture conditions, can differentiate into polarized enterocyte-like cells, making them a preferred model for intestinal epithelial barrier function, drug transport, and colorectal cancer signaling. Their endogenous expression of key components of the cAMP, Wnt, and Rho GTPase pathways renders them particularly suitable for studying the integrative role of AKAP13 in these networks.
AKAP13 (A-kinase anchoring protein 13, also known as AKAP-Lbc) is a large scaffold protein possessing both PKA-anchoring and RhoGEF domains. It binds PKA regulatory subunits to localize cAMP/PKA signaling to specific subcellular compartments, while its RhoGEF domain specifically activates RhoA. This dual activity enables AKAP13 to coordinate cAMP-dependent regulation of the actin cytoskeleton through the RhoA-ROCK-mDia pathway and to modulate serum response factor (SRF/MRTF-A)-mediated transcription. Furthermore, AKAP13 interacts with ??-catenin to influence Wnt/TCF/LEF target gene expression. Upstream, AKAP13 responds to G??s-coupled receptors such as the ??-adrenergic receptor and prostaglandin E2 receptor, linking extracellular signals to RhoA activation and gene transcription via adenylyl cyclase and cAMP.
In HT29 colorectal adenocarcinoma cells, AKAP13 integrates cAMP/PKA and Rho GTPase signaling to control actin dynamics, cell migration, and transcriptional programs relevant to tumor progression and intestinal epithelial homeostasis. Disruption of AKAP13 in this genetic background is expected to impair PKA anchoring, thereby perturbing localized cAMP responses, and to reduce RhoA activation, leading to defects in stress fiber formation, cell motility, and potentially in the Wnt/??-catenin-dependent transcription that drives proliferation and differentiation. Consequently, this knockout model offers a powerful platform to dissect the crosstalk between cAMP compartments and Rho-dependent cytoskeletal rearrangements in colorectal cancer cells.
This AKAP13 knockout polyclonal cell model is suitable for a wide range of biomedical research applications, including mechanistic studies of colorectal cancer signaling, analysis of intestinal epithelial cell migration and barrier function, and drug screening aimed at Rho/ROCK inhibitors or cAMP compartmentalization modulators. Researchers can employ assays such as RhoA activation (G-LISA), wound healing migration assays, immunofluorescence for filamentous actin, TCF/LEF luciferase reporter assays, and cAMP ELISA to characterize the functional consequences of AKAP13 loss. Additionally, transcriptomic profiling via RNA-seq and proliferation assays further enable comprehensive pathway analysis. For further technical details or ordering information, please contact Ascent Research.
