The AK7 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HEK293T human embryonic kidney cell line. This product comprises a heterogeneous mixture of cells harboring targeted disruptions in the AK7 gene, resulting in loss of adenylate kinase 7 function. The polyclonal format provides a diverse pool of knockout genotypes, enabling robust functional studies without the limitations of a single clonal isolate. By employing CRISPR/Cas9-mediated gene disruption, this model offers a reliable platform for investigating AK7-dependent biological processes.
HEK293T cells are a widely used derivative of the HEK293 line, stably expressing the SV40 large T antigen, which facilitates episomal replication of plasmids containing the SV40 origin of replication. Originating from female fetal human kidney, these cells are a common workhorse for protein expression, viral packaging, and diverse cell biology applications. Their robust growth and high transfection efficiency make them an ideal host for creating gene knockouts and studying fundamental cellular mechanisms.
Adenylate kinase 7 (AK7) is a key enzyme in purine metabolism, catalyzing the reversible interconversion of adenine nucleotides??ATP, ADP, and AMP. By precisely regulating intracellular adenine nucleotide pools and the AMP/ATP ratio, AK7 acts as a critical node linking energy homeostasis to downstream signaling. AK7 is regulated by transcription factors FOXJ1 and RFX family members and by energy stress. It modulates the activity of AMPK and PKA, and influences purinergic receptor responsiveness and ciliary beat frequency. AK7 interacts with other adenylate kinase isoforms such as AK8 and AK9, as well as with tubulins, underscoring its role in ciliary infrastructure. Mechanistically, AK7 governs the balance of adenine nucleotides, thereby tuning metabolic sensors like AMPK and the cAMP/PKA/CREB axis. In ciliated cells, this function is essential for maintaining proper ciliary motility, linking nucleotide metabolism directly to ciliogenesis and ciliary function.
In the HEK293T background, disruption of AK7 provides a powerful model to dissect the crosstalk between nucleotide metabolism and ciliary biology. Although HEK293T cells are not typically ciliated under standard culture conditions, they retain the molecular machinery for ciliogenesis and can be induced to form primary cilia. Thus, AK7 knockout HEK293T cells allow researchers to investigate how adenylate kinase activity influences ciliary assembly, maintenance, and signal transduction in a tractable cell-based system. This model is particularly relevant for studying ciliopathies such as primary ciliary dyskinesia and male infertility, where AK7 dysfunction has been implicated. It also facilitates examination of the gene??s role in energy stress responses and purinergic signaling pathways.
These polyclonal knockout cells are suitable for a range of functional assays, including Western blotting to confirm AK7 loss, RT-qPCR analysis of ciliary gene expression (e.g., FOXJ1 targets), bioluminescent quantification of ATP/AMP ratios, and immunofluorescence detection of cilia using acetylated ??-tubulin. They enable mechanistic studies of nucleotide homeostasis, drug target screening for ciliopathies, and evaluation of energy stress modulators. For further information or to discuss custom applications, please contact Ascent Research.
