The AKAP8 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population featuring targeted disruption of the AKAP8 gene in HeLa cells. This loss-of-function model is generated via CRISPR/Cas9-mediated gene disruption, resulting in a heterogeneous knockout pool that preserves population-level diversity. The polyclonal format avoids clonal selection artifacts and is ideal for functional studies requiring biological variability and robust gene-editing outcomes.
HeLa cells are an established human cervical adenocarcinoma line derived from Henrietta Lacks, characterized by epithelial morphology and stable HPV-18 positivity. Widely used in cancer and cell cycle research, HeLa offers well-defined genetics, ease of manipulation, and rapid growth, making it a standard host for CRISPR-mediated gene knockouts. The line’s aberrant cell cycle checkpoints and viral oncogene expression provide a relevant background for studying nuclear signaling and chromatin dynamics.
AKAP8 is a scaffold protein that anchors PKA to the nuclear matrix, directing cAMP-dependent phosphorylation to specific subnuclear sites. It interacts with PKA regulatory subunits RII?? and RII?? and is regulated upstream by cAMP and CDK1/cyclin B. Downstream, AKAP8 facilitates PKA-mediated phosphorylation of targets such as the SF2/ASF splicing factor and chromatin condensation proteins. It also associates with lamin B, topoisomerase II, and RNA helicase A, linking it to nuclear architecture and RNA processing. Key pathway members include adenylyl cyclase, cAMP, PKA catalytic and regulatory subunits, and AKAP8.
In HeLa cervical carcinoma cells, AKAP8 disruption allows interrogation of its role in cell cycle progression, mitotic chromosome condensation, and pre-mRNA splicing. Given the HPV-driven dysregulation of these pathways, AKAP8 knockout can uncover whether nuclear PKA anchoring is essential for proliferation or survival in a cancer-relevant setting. This model enables dissection of AKAP8-dependent spatial signaling in the context of oncogenic transformation.
Typical applications include cell cycle analysis via flow cytometry, immunofluorescence to track PKA localization, RT-qPCR to measure splicing isoform changes, and colony formation assays for proliferative assessment. Western blotting validates AKAP8 loss and downstream phosphorylation events. The knockout cells are also suited for cAMP/PKA signaling studies and mitotic dynamics. For more information, please contact Ascent Research.
