The ITFG1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the ITFG1 gene has been disrupted to generate a loss-of-function model. This heterogeneous pool of edited alleles avoids clonal biases, providing a reliable tool for studying ITFG1-dependent processes in adhesion, migration, and signaling. The product is supplied as ready-to-use near-haploid HAP1 cells, suitable for immediate expansion and downstream assays.
HAP1 is a near-haploid human fibroblast-like cell line originally derived from a chronic myeloid leukemia (CML) patient. Its stable haploid genome has established it as a premier system for functional genomics, enabling unambiguous interpretation of gene perturbations because a single functional allele is disrupted without compensatory wild-type expression. The cells retain key characteristics of mesenchymal lineages, including expression of integrins and focal adhesion components, making them appropriate for investigating adhesion dynamics.
ITFG1 encodes a transmembrane adaptor that couples integrin engagement to intracellular signaling. It binds integrins ??L??2 and ??4??1, recruiting FAK and Src to activate PI3K/AKT and MAPK pathways. ITFG1 also interacts with talin, vinculin, and LNX1, linking integrins to the actin cytoskeleton. Its activity is stimulated by TCR signaling, chemokines, and IL-2, positioning ITFG1 at the intersection of adhesion, migration, and immune activation.
Knocking out ITFG1 in the HAP1 background generates a clean loss-of-function system wherein the haploid state eliminates residual gene expression, enhancing the sensitivity of phenotypic readouts. This makes the model particularly valuable for unbiased synthetic lethality screens designed to identify genetic interactors that become essential upon ITFG1 loss. Moreover, HAP1 cells support immune-relevant signaling pathways, enabling investigation of ITFG1??s role in processes analogous to T cell activation and trafficking.
Applications include cell adhesion and spreading assays on ICAM?1 or fibronectin, transwell migration, and high?throughput CRISPR phenotypic screening. Biochemical validation via western blotting, co-immunoprecipitation, and immunofluorescence for focal adhesion proteins (paxillin, vinculin) is readily performed. Flow cytometry enables quantitative analysis of integrin surface expression and activation. For further details and technical support, please contact Ascent Research.
