The ITSN1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the near-haploid HAP1 cell line, designed for loss-of-function studies of the ITSN1 gene. This heterogeneous knockout pool allows researchers to interrogate ITSN1-dependent cellular processes without clonal selection artifacts. The product is provided as a proliferating cell culture suitable for both acute assays and long-term experimental regimes.
HAP1 is a near-haploid human cell line established from a chronic myeloid leukemia patient in blast crisis. With a haploid karyotype for most chromosomes, HAP1 offers a simplified genetic landscape that facilitates unambiguous genotype-phenotype associations. It is extensively used as a model system for cancer biology, particularly in genetic screens and investigations of leukemic signaling pathways.
ITSN1 encodes an evolutionarily conserved scaffold protein that couples clathrin-mediated endocytosis to intracellular signal transduction. It interacts with Dynamin and Synaptojanin via its EH and SH3 domains, while its DH-PH and SH3 regions engage SOS1, GRB2, and CBL to modulate downstream pathways. ITSN1 operates downstream of receptor tyrosine kinases such as EGFR and TrkA, and is activated by RAS and calcium influx. It promotes RAS-RAF-MEK-ERK cascade signaling, RHOA/CDC42 activation, and PI3K-AKT signaling. Disruption of ITSN1 therefore impairs clathrin-dependent internalization of growth factor receptors and attenuates critical MAP kinase and Rho family GTPase signaling outputs.
In the HAP1 background, ITSN1 knockout provides a clean genetic setting to examine scaffolding functions in a leukemic context. The absence of a second allele in this near-haploid line ensures that the loss-of-function phenotype is directly attributable to the edited locus, eliminating concerns of allelic compensation. This model enables dissection of endocytic trafficking and signaling networks that may contribute to CML pathogenesis, and supports identification of vulnerabilities arising from ITSN1 deficiency in cancer cells.
The ITSN1 Knockout HAP1 Polyclonal Cells support diverse applications, including transferrin uptake assays to measure clathrin-mediated endocytosis efficiency, biochemical analysis of phospho-ERK levels, and co-immunoprecipitation of endocytic complexes. Immunofluorescence studies can visualize altered clathrin-coated pit dynamics, while functional assays such as transwell migration and Rho GTPase activation pull-downs reveal downstream consequences of ITSN1 loss. Researchers in oncology, neurobiology, and signal transduction will find these cells valuable for target validation, drug response profiling, and mechanistic dissection of scaffold protein networks. For inquiries, contact Ascent Research.
