The AKAP17A Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted population of HeLa cells lacking functional AKAP17A. This polyclonal knockout model provides a heterogeneous loss-of-function background to study AKAP17A??s role in pre-mRNA splicing and cAMP/PKA signaling. Disruption of AKAP17A enables investigation of its scaffold function in anchoring protein kinase A to nuclear speckles for phosphorylation of splicing factors.
HeLa cells, derived from cervical adenocarcinoma, are immortalized epithelial cells containing HPV18 DNA with p53 and Rb inactivated by E6 and E7. Their hypertriploid karyotype and robust growth characteristics make them a widely used model for cancer research and gene expression studies. In this knockout context, the HeLa background provides a relevant tumorigenic environment for studying splicing dysregulation.
AKAP17A scaffolds PKA at nuclear speckles, coupling cAMP signals to RNA processing. cAMP-activated PKA catalytic subunits phosphorylate SR proteins such as SRSF1 and SRSF2 and other spliceosomal components, modulating alternative splicing of target pre-mRNAs. AKAP17A interacts with PKA regulatory subunits, U2AF, and SF1, and is regulated by cAMP and kinases SRPK1 and CLK1, thereby integrating extracellular cues with post-transcriptional gene control.
In the HeLa cervical cancer background, where splicing aberrations are common, AKAP17A knockout allows dissection of cAMP-driven splicing events that may contribute to oncogenic phenotypes. The loss of AKAP17A-mediated PKA anchoring can alter phosphorylation of SR proteins, shifting alternative splice patterns of genes involved in cell proliferation, apoptosis, or migration. With HeLa??s p53- and Rb-deficient status, this model isolates AKAP17A-dependent splicing contributions to cancer cell fitness without confounding tumor suppressor pathways.
This polyclonal knockout pool supports RT-PCR and RNA-seq for alternative splicing profiling, western blotting for phospho-PKA substrates, and immunofluorescence for nuclear speckle organization. It enables screening of splicing or cAMP modulators, and cell viability or apoptosis assays to assess AKAP17A??s impact on cancer cell survival. Co-immunoprecipitation studies can further define AKAP17A??s spliceosomal interactions. For further details, contact Ascent Research.
