The AKAP1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population targeting human AKAP1 in the HT29 colorectal adenocarcinoma line. This product provides a heterogeneous pool of cells with CRISPR-mediated gene disruption, enabling loss-of-function studies without clonal isolation. The polyclonal format is ideal for population-based assays and screening applications where averaging across genetic variants is acceptable.
HT29 cells originated from a primary colorectal adenocarcinoma of a 44-year-old female Caucasian and serve as a model of intestinal epithelium. They retain tumorigenic properties and can differentiate into enterocyte-like cells under defined conditions, making them suitable for research on colorectal cancer biology, differentiation, nutrient absorption, and drug permeability. The epithelial context is particularly relevant for investigating mitochondrial function in transformed intestinal cells.
AKAP1 acts as a mitochondrial scaffold that anchors PKA holoenzymes via regulatory subunits RII?? and RII?? to the outer membrane, thereby directing local cAMP signaling. This compartmentalization enables PKA-mediated phosphorylation of mitochondrial substrates, including DRP1 at Ser637 (inhibiting mitochondrial fission) and BAD at Ser112 (promoting cell survival). AKAP1 also targets COXIV-1 to modulate oxidative phosphorylation. Upstream regulators such as PPARGC1A, NRF1, and TFAM control AKAP1 expression, while interacting partners like PDE7A and PP1 fine-tune cAMP levels and dephosphorylation events. Src kinase additionally modifies AKAP1 function, integrating growth factor signals with mitochondrial dynamics and apoptosis.
In HT29 colorectal cancer cells, AKAP1-dependent mitochondrial PKA signaling influences apoptosis resistance, metabolic reprogramming, and chemosensitivity. By phosphorylating DRP1 and BAD, AKAP1 tilts the balance toward mitochondrial fusion and cell survival, potentially contributing to drug resistance in colorectal tumors. Disruption of AKAP1 in this model allows researchers to directly examine how local cAMP signaling impacts mitochondrial physiology, tumor cell viability, and response to therapies targeting the PKA pathway or mitochondrial machinery.
Researchers can employ these polyclonal knockout cells for diverse assays, including immunoblotting to monitor PKA substrate phosphorylation, RT-qPCR to assess transcriptional adaptations, and immunofluorescence to visualize mitochondrial network changes. Apoptosis and metabolic flux analyses (e.g., Seahorse) provide functional readouts, while migration and drug sensitivity assays reveal phenotypic consequences. The model supports screening for mitochondria-targeted compounds and mechanistic studies of AKAP1 in colorectal cancer metabolism and chemoresistance. For further technical inquiries, contact Ascent Research.
