The AKAP1 knockout HAP1 polyclonal cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HAP1 human cell line, carrying a targeted disruption of the AKAP1 gene. This loss-of-function model is produced through CRISPR/Cas9-mediated gene disruption, yielding a heterogeneous pool of cells with abrogated AKAP1 protein expression. The polyclonal format avoids clonal artifacts and provides a genetically diverse population for robust phenotypic assays and functional genomics screens.
HAP1 is a near-haploid human chronic myeloid leukemia cell line derived from the KBM-7 line, characterized by the BCR-ABL1 fusion oncogene and an adherent, fibroblast-like morphology. The near-haploid karyotype (one copy of most chromosomes) facilitates efficient CRISPR/Cas9 knockout generation and minimizes genetic redundancy, making HAP1 a widely adopted model for genetic screens and haploid genetic approaches. Its leukemic origin provides a relevant background for investigating oncogenic signaling, mitochondrial biology, and drug response mechanisms.
AKAP1 encodes a mitochondrial A-kinase anchoring protein that scaffolds protein kinase A (PKA) regulatory subunits (PRKAR1A and PRKAR2A) at the outer mitochondrial membrane. This localization mediates cAMP-dependent phosphorylation of critical substrates, including DNM1L (Drp1) at Ser637 to inhibit mitochondrial fission and BAD at Ser155 to suppress apoptosis. AKAP1 also interacts with VDAC1, SRC, MFN2, and OPA1, integrating signals from cAMP, calcium, AMPK, and mTOR pathways to regulate mitochondrial dynamics, intrinsic apoptosis, and cellular metabolism. Its disruption uncouples PKA from mitochondrial targets, altering cytochrome c release and caspase-9 activation.
In the HAP1 leukemic context, AKAP1 knockout is expected to disturb the spatial compartmentalization of cAMP/PKA signaling at mitochondria, thereby shifting the balance of mitochondrial fission/fusion, enhancing apoptotic priming, and impairing metabolic flexibility that supports BCR-ABL1-driven proliferation. The near-haploid genome simplifies identification of genetic interactions and facilitates synthetic lethality screens with drugs such as tyrosine kinase inhibitors or Bcl-2 antagonists. This model thereby serves as a powerful tool for dissecting mitochondrial contributions to leukemia cell fitness and treatment resistance.
The AKAP1 knockout HAP1 polyclonal cells support a wide range of experimental approaches. Biochemical analyses include Western blotting for phospho-Drp1 (Ser637) and total Drp1, co-immunoprecipitation of PKA subunits with AKAP1, and phospho-signaling arrays. Functional studies encompass immunofluorescence staining with MitoTracker to visualize mitochondrial morphology, flow cytometric apoptosis assays (Annexin V, cleaved caspase-3), and Seahorse metabolic flux analysis to measure oxygen consumption and glycolysis. Drug sensitivity profiling with Bcl-2 inhibitors (e.g., venetoclax) or BCR-ABL1 inhibitors can reveal AKAP1-dependent vulnerabilities. The polyclonal population is also well-suited for pooled CRISPR screens and long-term adaptation studies. For additional information, please contact Ascent Research.
