The CASS4 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the CASS4 gene has been disrupted using CRISPR/Cas9-mediated gene targeting. This product provides a heterogeneous knockout model suitable for functional studies of CASS4 in human near-haploid cells. As a polyclonal population, the cells can be used for loss-of-function experiments such as signaling assays, drug response testing, and genetic screening without requiring clonal isolation.
The host cell line, HAP1, is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) cell line, with a male origin and a haploid karyotype. HAP1 cells serve as a robust model for functional genomics and CRISPR-based knockout studies due to their single gene copy number, which simplifies genotype-phenotype correlations. They maintain key features of CML cells, including expression of the BCR-ABL fusion kinase, making them valuable for studying oncogenic signaling and drug responses in a haploid background.
CASS4 (Cas scaffolding protein family member 4) is a scaffolding protein critical for integrin-mediated signaling, cell adhesion, and migration. It is a known substrate of Src family kinases and interacts with focal adhesion kinase (FAK), p130Cas (BCAR1), and the adaptor protein Crk. CASS4 operates downstream of integrin activation and BCR-ABL kinase, coupling adhesive signals to the MAPK and PI3K/Akt pathways. By organizing focal adhesion complexes, CASS4 modulates cytoskeletal dynamics and cell motility, contributing to processes relevant to cancer invasion and metastasis.
Knockout of CASS4 in HAP1 cells disrupts integrin-dependent signaling networks, leading to impaired cell adhesion and migration. Given the haploid nature of HAP1 cells, disruption of a single CASS4 allele is sufficient to create a complete loss-of-function model. This system enables precise investigation of CASS4’s role in BCR-ABL-mediated signaling and its interplay with Src kinases and focal adhesion proteins. The loss of CASS4 may alter the phosphorylation status of FAK and p130Cas, providing a platform to study Src/FAK signaling cascades. Moreover, in the context of CML, this model can be used to evaluate the contribution of CASS4 to leukemic cell survival and migration, as well as to test sensitivity to tyrosine kinase inhibitors like imatinib.
The CASS4 Knockout HAP1 Polyclonal Cells are suitable for a wide range of functional assays, including western blotting, RT-qPCR, and Sanger sequencing to confirm knockout and assess downstream effects. Cell-based assays such as adhesion assays, Transwell migration assays, and colony formation assays can be employed to quantify phenotypic changes. Phospho-FAK and phospho-Akt western blotting can be used to evaluate signaling pathway disruptions. Additionally, these cells can be integrated into CRISPR-based genetic screens or used for drug target validation in CML and other cancer models. For further information on experimental use or customization, please contact Ascent Research.
