The IGSF8 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal cell population engineered to disrupt the human IGSF8 gene within the HAP1 host cell background. This product provides a mixed pool of edited cells, each carrying a targeted gene disruption generated by CRISPR/Cas9 technology, resulting in a loss-of-function model for studying IGSF8 biology. The polyclonal format is particularly suited for pooled functional screens and assays where heterogeneous knockout events are analytically advantageous.
HAP1 is a near-haploid human myeloid cell line derived from a male patient with chronic myeloid leukemia in blast crisis. As a suspension-adapted subline of KBM-7, HAP1 cells maintain a near-haploid karyotype with the exception of chromosome 8, which remains disomic. This haploid configuration simplifies genetic manipulation, enabling efficient generation of knockout models without the confounding effects of multiple gene copies, and has established HAP1 as a widely used platform for genome-wide knockout screens and pathway dissection.
IGSF8 encodes a cell surface immunoglobulin superfamily member that negatively regulates cell migration and invasion. It forms complexes with tetraspanins CD9 and CD81, integrating into the tetraspanin web to modulate integrin-mediated signaling. IGSF8 activity is regulated upstream by cell adhesion, integrin engagement, and TGFB1, while downstream it controls the phosphorylation of FAK and SRC kinases, and the activity of Rho GTPases RAC1 and RHOA, as well as AKT. These protein complexes coordinate integrin alpha3beta1 and alpha6beta1, influencing cytoskeletal dynamics and cell-matrix interactions.
In the near-haploid HAP1 background, IGSF8 knockout removes a negative regulator of migration, enabling dissection of its role in cell adhesion and signaling. The absence of additional alleles ensures high knockout efficiency and a robust loss-of-function phenotype. This model is relevant to cancer metastasis, as IGSF8 dysregulation occurs in melanoma, breast, and prostate cancers. The polyclonal knockout population allows functional studies on how IGSF8 deficiency alters integrin-driven signaling and cytoskeletal reorganization.
Researchers can employ these polyclonal knockout cells in transwell migration and invasion assays, cell adhesion assays, and western blotting for FAK and SRC phosphorylation. Flow cytometry confirms IGSF8 loss, while co-immunoprecipitation with CD9 and CD81 examines tetraspanin complex integrity. RT-qPCR for migration-related genes and drug sensitivity profiling are also feasible. The polyclonal pool is amenable to haploid genetic screens. For further details, please contact Ascent Research.
