The JUP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human JUP gene in HAP1 cells. This loss-of-function model enables researchers to investigate the consequences of plakoglobin deficiency in a genetically tractable background. The polyclonal format provides a heterogeneous pool of edited alleles, reflecting the range of functional disruptions achievable through CRISPR/Cas9-mediated gene targeting.
HAP1 is a near-haploid human chronic myeloid leukemia (CML) cell line derived from the KBM-7 line. Its near-haploid karyotype simplifies genetic screening and knockout studies, as single-copy gene disruptions generate unambiguous phenotypes without confounding from second alleles. The BCR-ABL positive background is typical of CML models, and the line retains key cellular machinery for adhesion and signaling research.
JUP encodes plakoglobin (??-catenin), a bifunctional protein that serves as a structural component of desmosomes and adherens junctions and as a transcriptional co-regulator in Wnt signaling. Plakoglobin interacts with E-cadherin (CDH1), desmoglein-2, desmocollin-2, ??-catenin, ??-catenin, desmoplakin, and plakophilin-2 to maintain cell-cell adhesion. In the Wnt pathway, plakoglobin, along with ??-catenin, is regulated by a destruction complex containing GSK3B, APC, and Axin. Wnt activation via WNT3A stabilizes plakoglobin, allowing its nuclear accumulation where it partners with TCF/LEF transcription factors such as TCF7L2 to modulate expression of target genes like MYC, CCND1, and DSC2. Thus, JUP knockout disrupts both junctional integrity and canonical Wnt transcriptional output.
In HAP1 cells, knockout of JUP generates a controlled system to dissect the role of plakoglobin in adhesion and signaling pathways. The near-haploid background ensures that loss-of-function phenotypes are not masked by redundant alleles, facilitating clear interpretation of effects on junction assembly and Wnt-mediated transcription. This model is particularly useful for studying diseases such as arrhythmogenic right ventricular cardiomyopathy (ARVC) and Naxos disease, where desmosomal dysfunction is central. Additionally, it aids in cancer research, where altered adhesion and Wnt signaling contribute to tumor progression and metastasis.
Researchers can employ these polyclonal knockout cells in a variety of applications, including quantitative Western blotting and immunofluorescence to assess the expression and localization of desmosomal and adherens junction markers such as plakoglobin, E-cadherin, and desmocollin-2. Cell adhesion aggregation assays and Wnt luciferase reporter assays provide functional readouts of adhesion integrity and transcriptional activity. Furthermore, co-immunoprecipitation of desmosomal complexes and transwell migration/invasion assays allow detailed mechanistic studies. This product is ideally suited for drug response screening in adhesion-deficient models and investigations into cancer cell migration and invasion. For additional information, please contact Ascent Research.
