The DIRAS1 Knockout HAP1 Polyclonal Cells product is a CRISPR/Cas9-edited polyclonal knockout cell population in which the tumor suppressor gene DIRAS1 has been disrupted to generate a loss-of-function model. This polyclonal format provides a heterogeneous cell population carrying a variety of CRISPR-induced edits at the target locus, enabling robust gene perturbation studies without the selection of a single clone. The knockout is achieved in the HAP1 host cell line, a near-haploid human cell model. This product is supplied as a live cell population and is intended for advanced biomedical research applications in cancer biology, signal transduction, and functional genomics.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia line and exhibit a fibroblast-like morphology with a near-haploid karyotype. This male-derived cell line carries the Philadelphia chromosome, resulting in BCR-ABL1 fusion expression. The near-haploid genome simplifies recessive genetic screens, as the presence of a single allele per gene facilitates the generation and analysis of knockout phenotypes. This characteristic makes HAP1 an excellent host for CRISPR-based gene disruption, enabling the functional interrogation of tumor suppressors like DIRAS1 with minimal confounding effects from wild-type alleles.
DIRAS1 encodes a small GTPase that functions as a tumor suppressor by negatively regulating cell proliferation and survival. Its expression is controlled by upstream regulators such as E2F1 and E2F4 transcription factors, as well as epigenetic mechanisms including DNA methylation and histone modifications. Mechanistically, DIRAS1 antagonizes RAS signaling and downregulates the PI3K/AKT and JAK/STAT3 pathways. This leads to reduced phosphorylation of AKT and ERK1/2, diminished STAT3 transcriptional activity, and subsequent induction of apoptosis and autophagy. Key downstream effectors include BCL2 family members (BCL2, BAX) and autophagy regulators (LC3, Beclin-1). DIRAS1 directly interacts with RAS proteins and nucleotides GTP/GDP to exert its suppressive effects.
In the near-haploid HAP1 background, disruption of DIRAS1 is expected to relieve its tumor-suppressive constraints, resulting in enhanced proliferation, survival, and potentially impaired autophagy and apoptosis. This model thus provides a defined cellular context to study the molecular consequences of DIRAS1 loss, which is associated with malignancies such as ovarian, breast, thyroid, pancreatic cancers, and glioblastoma. Because HAP1 cells retain key signaling pathways, including RAS and PI3K/AKT, the knockout population enables the examination of pathway crosstalk and the identification of synthetic lethal interactions.
This product is suited for a broad spectrum of research applications, including the dissection of tumor suppressor mechanisms, drug target discovery for cancers exhibiting DIRAS1 silencing, and epigenetic therapy studies aimed at restoring DIRAS1 expression. Representative experimental readouts include western blotting for AKT and STAT3 phosphorylation, cell proliferation assays (MTS, BrdU), apoptosis assays (annexin V, caspase activation), autophagy flux measurements (LC3-II turnover), RT-qPCR for downstream target genes, colony formation assays, and migration/invasion tests. Co-immunoprecipitation assays can be employed to probe DIRAS1?CRAS interactions. For further information or to discuss your specific project needs, please contact Ascent Research.
