This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human AKAP8L gene in HEK293T epithelial cells. The cells bear heterogeneous loss-of-function mutations introduced by CRISPR/Cas9, providing a reliable model for investigating AKAP8L-dependent processes without clonal isolation. The polyclonal format preserves genetic diversity, enabling consistent knockout phenotypes while reducing clonal artifacts.
HEK293T cells are derived from human embryonic kidney cells transformed with adenovirus type 5 DNA and stably expressing SV40 large T antigen. This genetic background supports high-level episomal plasmid amplification and robust protein expression, making the cells a preferred host for studies on nuclear dynamics, signal transduction, and viral vector production. Their epithelial origin and technical tractability facilitate gene-editing applications.
AKAP8L (A-kinase anchoring protein 8-like) acts as a scaffold that tethers PKA regulatory subunits to the inner nuclear membrane through direct binding to LAP2beta and emerin. This positioning brings PKA into proximity with lamin A/C, enabling phosphorylation events essential for mitotic nuclear envelope breakdown and reassembly. Upstream, CDK1/cyclin B and cAMP-regulated PKA control AKAP8L-mediated signaling; downstream, the pathway governs chromosome segregation and lamin dynamics. AKAP8L also integrates with RNA processing machinery, interacting with Btf and RNA polymerase II to modulate pre-mRNA splicing and transcription. Consequently, AKAP8L dysfunction has been implicated in chromosomal instability, nuclear envelopathies, and developmental abnormalities.
Within the HEK293T context, AKAP8L knockout disrupts a critical signaling node at the nuclear envelope, directly affecting mitosis and gene expression regulation. The polyclonal knockout population allows bulk analysis of nuclear lamina phosphorylation, cell cycle progression, and splicing fidelity without the need for clonal selection. This format is well-suited for population-based assays such as flow cytometry, live-cell imaging, and high-throughput screens, where reproducible, averaged phenotypes are desired over clonal variation.
This AKAP8L knockout model supports a range of applications. Co-immunoprecipitation and Western blotting can probe AKAP8L?CPKA complex stability and phospho-lamin A/C levels; immunofluorescence microscopy visualizes nuclear envelope architecture and protein localization. RNA-seq and splicing-sensitive PCR enable assessment of transcriptional and post-transcriptional changes. The model is particularly relevant for oncology research on mitotic catastrophe and for exploring pathomechanisms of laminopathies. For additional details, please contact Ascent Research.
