The KCTD12 Knockout HAP1 Polyclonal Cells product comprises a pool of CRISPR/Cas9-edited HAP1 cells with targeted disruption of the KCTD12 locus. This polyclonal knockout population provides a genetically heterogeneous loss-of-function model for functional studies without clonal isolation, enabling robust assessment of KCTD12-dependent phenotypes. In the HAP1 background, it facilitates investigation of KCTD12’s roles in receptor modulation and ubiquitin-mediated proteostasis.
The HAP1 cell line is a near-haploid human model derived from KBM-7 chronic myeloid leukemia cells, with a haploid karyotype disomic only for chromosome 8 and part of chromosome 15. This adherent line is widely used in haploid genetic screens, gene-trap mutagenesis, and CRISPR-based functional genomics because the single allele for most genes simplifies genotype-phenotype correlations. Its near-haploidy enhances genetic perturbation efficiency and reduces redundancy, making it an ideal knockout host.
KCTD12 encodes a dual-function protein: it serves as an auxiliary subunit of GABAB receptors, interacting with GABBR1 and GABBR2 to modulate cAMP signaling, and as a substrate adaptor for the Cullin3-RING E3 ubiquitin ligase complex, binding CUL3 to promote ubiquitination and degradation of targets such as ??-catenin. This links KCTD12 to Wnt/??-catenin pathway and apoptosis, with downstream upregulation of BAX and downregulation of BCL2. Transcriptionally regulated by SP1, CREB1, and miR-137, KCTD12 also cooperates with KCTD8 and KCTD16.
In the HAP1 near-haploid background, KCTD12 disruption allows direct examination of its tumor-suppressive and signaling functions. Loss of KCTD12-mediated ??-catenin ubiquitination stabilizes ??-catenin, enhancing Wnt pathway output and potentially altering proliferation and apoptosis. The polyclonal knockout pool mirrors heterogeneous responses in tissues and tumors, useful for studying variability in GABAB signaling and Cullin3-dependent degradation. The haploid architecture facilitates secondary CRISPR screens for synthetic lethal interactions or resistance mechanisms in cancers like gastrointestinal stromal tumors and colorectal cancer.
This product is suited for western blotting of KCTD12, GABAB subunits, ??-catenin, BAX, BCL2; co-immunoprecipitation of KCTD12-CUL3 complexes; cAMP assays; ubiquitination assays; cell proliferation (MTS/MTT); and apoptosis (caspase-3/7) measurements. Employ in haploid genetic screens to identify regulators of GABAB signaling or ubiquitin-mediated proteolysis, and for target validation in neurological disorders such as schizophrenia and epilepsy. For specifications or custom inquiries, contact Ascent Research.
