The AKAP8 Knockout HEK293T Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal population of HEK293T cells bearing targeted disruption of the AKAP8 gene. This loss-of-function model provides a genetically defined system to interrogate AKAP8-dependent nuclear signaling without the bias of single-cell cloning.
HEK293T cells are a female human embryonic kidney epithelial line transformed with adenovirus 5 DNA and stably expressing the SV40 large T antigen. These adherent cells are widely utilized for high-level protein expression and virus production due to their high transfectability and robust growth characteristics.
AKAP8 encodes an A-kinase anchoring protein that scaffolds PKA to the nuclear matrix via direct interaction with PKA regulatory subunits RI?? and RII??. It is integral to cAMP/PKA signaling and regulates chromatin remodeling, nuclear receptor signaling, and gene expression. Key downstream targets include the transcription factor CREB, histone H3, and the androgen and estrogen receptors, while upstream regulators such as cAMP, PKA, PKC, and Aurora B kinase modulate its function. AKAP8 also interacts with emerin, lamin A/C, and HDAC3 to integrate nuclear architecture with signaling. Gene disruption abolishes PKA anchoring, impairing phosphorylation of chromatin-associated proteins and transcription factors and thereby altering cAMP-dependent gene regulation and chromatin dynamics.
In HEK293T cells, which natively express the cAMP signaling machinery and nuclear matrix components, the AKAP8 polyclonal knockout population offers a robust system for studying nuclear PKA functions without clonal bias. This model is highly relevant for cancer research, particularly breast and prostate cancers, and for investigating cardiovascular and neurological disorders where AKAP8-mediated signaling is implicated.
Typical applications include Western blotting for phospho-CREB and phospho-histone H3, RT-qPCR for c-Fos and AREG, immunofluorescence for nuclear localization, ChIP-qPCR for histone modifications, and co-immunoprecipitation of AKAP8 interactors. CREB reporter assays and RNA-seq facilitate transcriptomic analysis, while cell proliferation assays assess functional outcomes. These tools support drug target validation and mechanistic studies. For further details, please contact Ascent Research.
