The KPTN Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for loss-of-function studies of the KPTN gene in a human haploid cell background. This product comprises a heterogeneous pool of HAP1 cells harboring targeted disruptions in the KPTN locus, providing a robust model for investigating mTORC2 signaling and actin cytoskeleton regulation. The polyclonal format enables researchers to work with a genetically diverse knockout population, minimizing clonal artifacts and facilitating functional genomic screens.
The host cell line, HAP1, is a near-haploid adherent cell line derived from KBM-7 chronic myeloid leukemia (CML) cells. HAP1 cells exhibit a fibroblast-like morphology and retain a haploid karyotype for most chromosomes, making them exceptionally useful for genetic knockout and screening applications. Their use is well-established in high-throughput functional genomics, drug sensitivity profiling, and mechanistic cell biology, as the single-copy genome allows for unambiguous gene disruption without the need for homozygous targeting.
KPTN encodes a subunit of the mechanistic target of rapamycin complex 2 (mTORC2), where it interacts with mTOR, RICTOR, SIN1, and PROTOR. Functionally, KPTN links growth factor signaling to actin filament remodeling by directly binding F-actin and facilitating mTORC2-mediated phosphorylation of AKT at Ser473. This phosphorylation event is a key node downstream of insulin/IGF-1 receptor and PI3K activation, integrating cytoskeletal dynamics with cell survival and proliferation pathways. Additionally, KPTN participates in Rho GTPase signaling and cofilin regulation, further modulating cell morphology and migration.
Disruption of KPTN in the HAP1 haploid context provides a powerful system to dissect mTORC2-dependent actin organization independent of compensatory alleles. The loss of KPTN function impairs AKT phosphorylation at Ser473 and alters F-actin structures, enabling precise interrogation of how mTORC2 coordinates cellular architecture with signaling cues. Given the association of KPTN mutations with macrocephaly-intellectual disability syndrome and autism spectrum disorder, this knockout model is highly relevant for neurodevelopmental disease research, particularly in studying aberrant actin dynamics and mTOR dysregulation.
Researchers can employ these polyclonal knockout cells in a variety of applications, including quantitative Western blot analysis of phospho-AKT (Ser473), immunofluorescence staining of F-actin, and cell migration or invasion assays. The HAP1 background further supports drug sensitivity screening and RNA-seq transcriptomic profiling to identify downstream targets affected by KPTN loss. High-throughput genetic screening is also feasible, leveraging the haploid genome for synthetic lethality or modifier screens. For detailed product specifications and ordering information, please contact Ascent Research.
