The HEG1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal population of HAP1 cells carrying a targeted disruption of the HEG1 gene. This loss-of-function model provides a stable system to study HEG1-dependent signaling without the limitations of transient knockdowns. The polyclonal format offers a broad representation of edits, enabling robust functional genomic analyses in a well-defined genetic context.
The parental HAP1 cell line is derived from the near-haploid chronic myeloid leukemia (CML) line KBM-7 and is BCR-ABL positive. Its near-haploid karyotype, with only one copy of most chromosomes, eliminates confounding effects from heterozygous mutations and facilitates straightforward genotype-phenotype correlation. HAP1 cells serve as a versatile host for haploid genetic screens and are widely used to investigate myeloid hematopoietic biology and oncogenic signaling. Their rapid growth and amenability to genetic manipulation make them an ideal platform for generating knockouts and conducting high-throughput assays.
HEG1 encodes a transmembrane receptor that mediates endothelial cell adhesion and junction stability through DLL4-dependent recruitment of KRIT1 and CCM2. This pathway integrates Notch and Hippo signaling by suppressing YAP/TAZ transcriptional activity, thereby controlling angiogenesis and proliferation. Upstream regulators such as VEGF and Notch intracellular domain, along with downstream effectors like beta-catenin and VE-cadherin, form a network involving PDCD10 and ITGB1. Thus, HEG1 acts as a molecular hub linking Notch-mediated lateral inhibition with Hippo-regulated growth control.
In the HAP1 near-haploid context, HEG1 disruption simplifies dissection of Notch-Hippo crosstalk relevant to cardiovascular development and disease. Mutations in the KRIT1-CCM2-PDCD10 complex, associated with cerebral cavernous malformations and congenital heart defects, converge on HEG1-mediated junctional maintenance. Polyclonal knockout cells allow monitoring of pathway perturbations such as altered YAP phosphorylation or KRIT1 localization. The haploid nature ensures tight genotype-phenotype coupling, facilitating quantitative signaling studies and drug screens.
These cells are compatible with diverse assays including Western blot for phospho-YAP and KRIT1, immunofluorescence for VE-cadherin and beta-catenin, and cell adhesion/spreading assays. Functional analyses such as endothelial tube formation, Notch reporter luciferase assays, and co-immunoprecipitation of HEG1-KRIT1 complexes provide mechanistic insights, while qPCR for HES1 and HEY1 measures Notch output. Applications span cardiovascular development, angiogenesis, CCM modeling, Notch-Hippo crosstalk analysis, and drug screening for vascular disorders. For additional information, please contact Ascent Research.
