The DIRAS2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, designed to ablate expression of the tumor suppressor DIRAS2 gene. This polyclonal pool contains a heterogeneous mixture of gene-disrupted alleles generated by CRISPR/Cas9-mediated targeting, enabling loss-of-function studies without clonal isolation. The product provides a robust model for investigating DIRAS2-dependent signaling and its role in cancer biology.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) line, which harbors the BCR-ABL fusion oncogene. Its near-haploid karyotype minimizes genetic redundancy, making it an ideal platform for functional genomics, CRISPR-based knockout screens, and mechanistic studies of signaling pathways. The cell line retains key features of myeloid leukemia cells, providing a disease-relevant context for tumor suppressor gene research.
DIRAS2 encodes a small GTPase that functions as a tumor suppressor by antagonizing oncogenic RAS-driven signaling. It directly interacts with SmgGDS (RAP1GDS1) to inhibit activation of HRAS and KRAS, thereby suppressing downstream effector cascades including the MAPK/ERK and PI3K/AKT pathways. DIRAS2 is transcriptionally regulated by TP53 and E2F1, and its expression is frequently silenced by DNA methylation in cancers. The protein promotes apoptosis by modulating the BCL2 family balance, upregulating pro-apoptotic BAX and downregulating anti-apoptotic BCL2, and it inhibits cell cycle progression through cyclin D1 suppression. Additionally, DIRAS2 reduces tumor cell invasion by downregulating MMP2, and its loss leads to hyperactivation of AKT and ERK1/2 phosphorylation, driving uncontrolled proliferation and survival.
In HAP1 cells, which express the BCR-ABL fusion kinase that constitutively activates RAS/MAPK and PI3K/AKT signaling, disruption of DIRAS2 removes a critical brake on these oncogenic pathways. This polyclonal knockout model is therefore expected to exhibit enhanced proliferative capacity, reduced apoptosis, and altered sensitivity to tyrosine kinase inhibitors, making it a powerful tool for studying tumor suppressor function in a CML background. The absence of DIRAS2 may also uncover compensatory mechanisms or synthetic lethal interactions relevant to leukemia therapy.
These DIRAS2 knockout HAP1 polyclonal cells are suitable for a wide range of research applications, including functional genomics screens to identify synthetic lethal partners, mechanistic studies of RAS-driven oncogenesis, and evaluation of drug responses in CML and solid tumor models. Typical assays include monitoring cell proliferation via growth curves and colony formation, assessing apoptosis by Annexin V staining and western blot analysis of cleaved caspase-3 and PARP, and measuring signaling pathway activity through phosphorylated ERK1/2 and AKT levels. The model can also be employed in xenograft tumor studies to examine tumor growth and metastasis in vivo, as well as in CRISPR-based synthetic lethality screens to uncover novel therapeutic targets. For further details, please contact Ascent Research.
