This product comprises a CRISPR/Cas9-edited polyclonal knockout cell population of the human HAP1 cell line, in which the CCBE1 gene has been disrupted. The polyclonal nature of the population ensures representation of diverse genetic backgrounds while maintaining consistent loss-of-function across the pool. This knockout model is designed for researchers investigating the molecular mechanisms of CCBE1 in lymphangiogenesis and related signaling pathways. The cells provide a robust tool for both targeted mechanistic studies and broader phenotypic screening applications.
The HAP1 host cell line is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia cell line. As a leukemic hematopoietic progenitor model, HAP1 cells exhibit a simplified genetic landscape that facilitates gene targeting and functional genomic analyses. Their near-haploid karyotype allows efficient CRISPR/Cas9-mediated gene disruption and downstream clonal studies. This background is particularly advantageous for examining the biochemical and cell-signaling roles of extracellular matrix proteins such as CCBE1 without the complexity of diploid genetic compensation.
CCBE1 is an extracellular matrix glycoprotein that critically enhances the ADAMTS3-mediated proteolytic cleavage of pro-VEGF-C, generating mature VEGF-C. The cleaved VEGF-C then binds to VEGFR-3, often in complex with the co-receptor NRP2, triggering receptor phosphorylation and downstream signaling cascades that drive lymphatic endothelial cell commitment and lymphangiogenesis. CCBE1 function is regulated by transcription factors including PROX1, SOX18, and COUP-TFII, and it operates upstream of VEGF-C/VEGFR-3 signaling. Mechanistically, CCBE1 interacts with ADAMTS3 and collagen, facilitating the processing of pro-VEGF-C within the extracellular matrix. Disruption of CCBE1 thus impairs lymphangiogenic signaling, making this knockout model invaluable for dissecting these molecular interactions.
Although HAP1 cells are leukemic in origin and do not recapitulate the lymphatic endothelial context, their genetic tractability makes them an ideal platform for studying CCBE1-mediated signaling in a reductionist system. This knockout population allows researchers to probe the direct biochemical functions of CCBE1, such as its role in ADAMTS3 activation and VEGF-C processing, in a controlled cellular environment. The model can be used to validate protein?Cprotein interactions, assess signaling kinetics, and screen for modulators of the CCBE1-ADAMTS3-VEGF-C axis without confounding factors present in primary lymphatic cells.
Key applications include investigating lymphatic development and disease modeling for Hennekam lymphangiectasia-lymphedema syndrome, screening for lymphangiogenesis modulators using VEGF-C cleavage assays or phospho-VEGFR-3 western blotting, and performing functional genomics studies on extracellular matrix remodeling. The knockout cells are suitable for lymphangiogenesis sprouting assays, cell migration assays, and co-immunoprecipitation to examine CCBE1-ADAMTS3 interactions. Researchers can leverage this model to explore both canonical and non-canonical CCBE1 signaling pathways. For further information, please contact Ascent Research.
