The HECA Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HAP1 near-haploid human cell line, engineered to disrupt the HECA gene. This product provides a loss-of-function model for investigating HECA??s role as a transcriptional co-repressor in Wnt, TGF-beta, and Notch signaling pathways. The polyclonal format offers a cost-effective and versatile tool for functional genomics studies, avoiding the need for single-cell clone isolation while maintaining a heterogeneous knockout pool suitable for pooled screening and population-level analyses.
The host cell line HAP1 is a near-haploid chronic myeloid leukemia cell line derived from KBM-7. Its haploid nature (except for a disomic chromosome 15 fragment) simplifies genetic manipulation as most genes are present in a single copy, increasing the probability of complete knockout. HAP1 cells are widely employed for CRISPR-based screens, knockout validation, and signaling studies due to their robust growth and ease of culture. The leukemia background also makes this model relevant for hematological malignancy research.
HECA encodes a transcriptional co-repressor that physically interacts with TCF/LEF (e.g., TCF7L2, LEF1) and SMAD (SMAD2/3/4) transcription factors, thereby repressing Wnt and TGF-beta target gene expression. It functions downstream of Wnt ligands through beta-catenin (CTNNB1) and TGF-beta receptor activation. HECA negatively regulates cell cycle progression by repressing pro-proliferative genes such as MYC and CCND1, while promoting expression of the cyclin-dependent kinase inhibitor CDKN1A. Through these interactions, HECA acts as a potential tumor suppressor, with dysregulation linked to head and neck squamous cell carcinoma and other cancers.
In the HAP1 near-haploid background, disruption of HECA eliminates its repressive function, leading to potential derepression of Wnt and TGF-beta target genes. This simplified genetic context allows clear dissection of HECA-dependent signaling events without interference from a second allele. The combination of a leukemia-derived host with targeted HECA knockout creates a powerful system to study the role of this co-repressor in cell cycle control, differentiation, and tumorigenesis, particularly in hematopoietic contexts.
These polyclonal knockout cells are suitable for a range of experimental applications, including Wnt/TGF-beta pathway analysis using TOP/FOP Flash luciferase reporters, cell cycle profiling by flow cytometry, and gene expression analysis via RT-qPCR or RNA-seq. They can be employed in genetic screens for synthetic lethality, drug sensitivity testing, and apoptosis assays. The polyclonal population is also amenable to western blotting and immunofluorescence for protein detection and localization studies. For further information or to discuss custom projects, please contact Ascent Research.
