The AKAP13 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid human HAP1 cell line. This product provides a loss-of-function model for AKAP13, a dual-function scaffold protein that coordinates PKA anchoring and RhoA guanine nucleotide exchange factor activity. The heterogeneous knockout pool enables population-level biochemical and functional analyses without the biases of clonal selection, making it well-suited for studies of AKAP13-dependent signaling.
HAP1 cells originate from a male chronic myeloid leukemia patient (KBM-7 subclone) and exhibit a near-haploid karyotype that simplifies genetic manipulation and reduces allelic complexity. Widely used in genetic screens, these cells retain functional Rho GTPase, GPCR, and integrin signaling pathways, providing a relevant background for dissecting AKAP13-mediated cytoskeletal regulation and signal transduction.
AKAP13 integrates cAMP/PKA and RhoA signaling through distinct structural domains. Its A-kinase binding domain anchors PKA regulatory subunits, directing targeted phosphorylation of substrates downstream of cAMP-generating GPCRs and other stimuli. The tandem Dbl homology domain acts as a RhoGEF, activating RhoA to stimulate ROCK, LIMK, and cofilin, thereby promoting actin stress fiber formation and cell contractility. Upstream regulators include integrins, growth factor receptors, and G??12/13-coupled receptors, while downstream effectors such as SRF, MRTF-A, YAP/TAZ, and ??-catenin transduce mechanical and chemical signals into transcriptional responses and cytoskeletal rearrangement.
Knockout of AKAP13 in HAP1 cells disrupts localized PKA activity and RhoA activation, leading to aberrant F-actin organization, impaired cell migration, and dysregulated SRF/YAP/TAZ-driven transcription. The haploid genetic background offers a streamlined system for interrogating AKAP13’s role in cancer-related processes like invasion and Rho-mediated mechanotransduction, as well as its involvement in Wnt and focal adhesion pathway crosstalk.
Representative assays for this product include Western blotting for AKAP13 and downstream phospho-targets, RhoA activation G-LISA, immunofluorescence staining of F-actin, Transwell migration and invasion assays, co-immunoprecipitation of PKA-AKAP13 complexes, phospho-PKA substrate profiling, and SRF/MRTF-A luciferase reporter assays. These techniques support investigations into cell motility, GPCR-to-RhoA signaling, PKA compartmentalization, and high-throughput screening for modulators of AKAP13 interactions. For further information, please contact Ascent Research.
