The AKAP7 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely utilized HEK293T human embryonic kidney epithelial cell line. In this product, the gene encoding A-kinase anchoring protein 7 (AKAP7) has been disrupted to generate a loss-of-function model that allows researchers to investigate the consequences of AKAP7 deficiency in a high-transfectability host. The polyclonal nature of the knockout population provides a heterogeneous mixture of edited cells, enabling robust and scalable experimental workflows without the clonal selection biases inherent in monoclonal lines.
The host cell line, HEK293T, is a derivative of HEK293 cells stably expressing the SV40 large T antigen. This modification enables episomal replication of plasmids containing the SV40 origin, yielding exceptionally high transfection efficiencies and protein expression levels. Originally derived from human embryonic kidney cells transformed with sheared adenovirus 5 DNA, HEK293T cells are an established model for heterologous expression, viral production, and cell signaling studies, making them an ideal chassis for examining AKAP7 function in a non-cardiac, non-neuronal background.
AKAP7 functions as a critical scaffold that anchors protein kinase A (PKA) holoenzymes to specific subcellular compartments, thereby confining cAMP/PKA signaling to discrete microdomains. Through its interaction with PKA regulatory subunits (RI??, RII??), AKAP7 orchestrates the phosphorylation of downstream targets including the L-type calcium channel (CACNA1C), KCNQ1 potassium channel, phospholamban (PLN), ryanodine receptor (RYR2), and AMPA receptor subunit GRIA1. This compartmentalized signaling is activated by ??-adrenergic stimulation and cAMP elevation, and it modulates processes ranging from cardiac muscle contraction and calcium handling to synaptic plasticity. Loss of AKAP7 disrupts these anchored complexes, leading to diffuse and aberrant phosphorylation patterns.
In the HEK293T background, AKAP7 knockout results in a loss of localized PKA signaling and a redistribution of kinase activity that can unmask non-canonical or compensatory signaling pathways. While HEK293T cells lack the full complement of cardiac- or neuron-specific ion channels, their robust protein expression machinery and amenability to heterologous reconstitution make them a powerful platform for dissecting AKAP7-dependent signaling events. Researchers can co-express relevant PKA substrates??such as wild-type or mutant CACNA1C, KCNQ1, or phospholamban??to examine how AKAP7 deficiency alters their phosphorylation, trafficking, and function in a controlled cellular environment. This model thus provides a versatile system for probing the mechanistic underpinnings of AKAP7-mediated signal compartmentation.
Typical applications include studying cAMP/PKA compartmentation using FRET-based cAMP sensors, assessing PKA substrate phosphorylation via western blotting, validating AKAP7?Cprotein interactions through co-immunoprecipitation, and screening for small-molecule disruptors of AKAP/PKA complexes. Additionally, the polyclonal knockout cells can be employed in electrophysiological recordings after transient transfection of ion channels or in calcium imaging experiments to investigate altered calcium dynamics. By eliminating endogenous AKAP7, this model enables clear interpretation of signaling perturbation experiments and accelerates the identification of AKAP7-dependent regulatory nodes. For further information or to place an order, please contact Ascent Research.
