The AKAP17A Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from HEK293T human embryonic kidney cells. This loss-of-function model targets AKAP17A, a gene encoding an A-kinase anchoring protein that scaffolds PKA to the spliceosome. The polyclonal format, generated through gene disruption without single-cell cloning, captures heterogeneous knockout alleles, allowing researchers to assess collective effects on alternative splicing and cAMP/PKA signaling.
HEK293T cells are a widely used derivative of HEK293, transformed with adenovirus 5 E1A/E1B and expressing SV40 large T antigen. These features confer high transfection efficiency and support episomal plasmid replication, making them ideal for protein overexpression and viral production. Their embryonic kidney epithelial origin provides a robust background for studying RNA processing and signal transduction. In this model, HEK293T serves as a versatile platform to investigate AKAP17A function.
AKAP17A anchors PKA to the spliceosome, where it phosphorylates serine/arginine-rich splicing factors SRSF1 and SRSF2, modulating their activity and regulating alternative splicing of targets like CD44 and CTNND1. Upstream, cAMP activates PKA; protein kinase C may influence localization. AKAP17A interacts with the PKA regulatory subunit PRKAR1A and spliceosome component SF3B1. Thus, it integrates cAMP/PKA signaling with spliceosome assembly, coupling signal transduction to post-transcriptional regulation.
Disrupting AKAP17A in HEK293T delocalizes PKA from the spliceosome, altering SRSF phosphorylation and splicing profiles. The high transfectability of HEK293T facilitates rescue experiments with wild-type or mutant AKAP17A, enabling structure-function studies. The polyclonal population avoids clonal bias and better reflects heterogeneous cellular responses, making it valuable for cancer and splicing-related disorder research where splicing dysregulation is implicated.
Researchers can utilize the AKAP17A Knockout HEK293T Polyclonal Cells to dissect alternative splicing mechanisms through assays such as RT-qPCR for splicing isoform quantification, Western blotting to detect phosphorylated SRSF levels, and RNA-seq for global splicing analysis. Co-immunoprecipitation experiments can probe interactions between AKAP17A, PKA, and spliceosome factors, while immunofluorescence microscopy localizes these proteins. These approaches enable comprehensive investigation of AKAP17A function in PKA signaling and RNA processing. For further technical information, please contact Ascent Research.
