AKAP9 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population in which AKAP9 has been disrupted, creating a loss-of-function model for studying A-kinase anchoring protein 9 in human colorectal adenocarcinoma. The heterogeneous pool of edited HT29 cells avoids clonal selection bias and is suitable for population-level analyses of centrosome biology, Golgi organization, and cAMP/PKA signaling.
The HT29 cell line, derived from a primary colorectal adenocarcinoma of a 44-year-old female, is an epithelial model capable of differentiating into enterocyte- and goblet-like phenotypes. This line provides a well-characterized background for investigating colorectal cancer cell cycle regulation, differentiation, and responses to cAMP pathway modulators.
AKAP9 encodes a scaffold that anchors PKA type II regulatory subunits (PKA-RII) to the centrosome and Golgi, compartmentalizing cAMP-dependent phosphorylation. It integrates signals from adenylate cyclase/cAMP/PKA and mitotic kinases Aurora A and CDK1?Ccyclin B. At the centrosome, AKAP9 assembles complexes with pericentrin, CDK5RAP2, CEP68, and ??-tubulin, and it controls the phosphorylation of downstream targets NDE1, NDEL1, and GM130. Consequently, AKAP9 regulates G2/M transition, centrosome maturation, microtubule organization, and Golgi integrity. Mutations in AKAP9 are associated with long QT syndrome, breast and colorectal cancers, and centriole-related ciliopathies.
Disruption of AKAP9 in HT29 cells is predicted to abolish PKA anchoring, leading to defects in mitotic progression, centrosome amplification, and Golgi apparatus disorganization. Because HT29 cells retain differentiation potential, the knockout may also impair enterocytic polarity and secretory functions, offering a model to parse the contributions of compartmentalized PKA signaling to colorectal cancer pathology and to test strategies targeting the AKAP9 scaffold.
These polyclonal knockout cells support diverse applications: cell cycle analysis by flow cytometry, immunofluorescence of centrosomal (pericentrin, ??-tubulin) and Golgi (GM130) markers, co-immunoprecipitation of PKA subunits, proliferation (MTS) and migration (Boyden chamber) assays, RNA-seq profiling, and small-molecule screening for AKAP9 interaction disruptors. For more details, contact Ascent Research.
