The ID1 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population engineered for ID1 gene disruption in the near-haploid human HAP1 cell line. This loss-of-function model leverages the haploid background, where single-copy gene targeting yields complete knockout phenotypes without allele compensation, facilitating functional genomics and genetic screens.
HAP1 is a near-haploid derivative of the KBM-7 chronic myeloid leukemia (CML) cell line, isolated from a male patient in blast crisis. The haploid karyotype simplifies gene-editing workflows, as disruption of a single allele suffices to ablate gene expression, thereby reducing variability inherent to heterozygous systems. HAP1 cells retain key signaling pathways of hematopoietic cancers, making them a relevant platform for leukemia research and drug sensitivity studies.
ID1 acts as a dominant-negative regulator of basic helix-loop-helix (bHLH) transcription factors, including TCF3 (E2A), TCF4, MYOD1, and TWIST1, by forming inactive heterodimers that impede DNA binding. Upstream, ID1 is transcriptionally induced by BMP2/BMP4 through BMPR1A-mediated SMAD1 phosphorylation and subsequent nuclear translocation with SMAD4, with further modulation by TGF-??1, EGF, and PDGF signaling. Knockout of ID1 relieves suppression of downstream targets such as CDKN1A (p21), CDKN2A (p16), VEGFA, MMP2, and BCL2, thereby derepressing cell cycle arrest, apoptosis, and angiogenic programs.
In the leukemic HAP1 context, loss of ID1 disrupts pro-proliferative and pro-survival signaling, partially through derepressed CDKN1A expression and altered BCL2 levels. This model is particularly suited for haploid genetic screens designed to uncover synthetic lethal interactions or drug targets that are selectively essential in ID1-deficient cells. The polyclonal population maintains background genetic heterogeneity, providing a more physiologically relevant system than monoclonal isolates.
Applicable assays include RT-qPCR and Western blotting to quantify ID1 and downstream targets, flow cytometry for cell cycle profiling, transwell migration and invasion assays, and luciferase-based bHLH reporter systems. This product supports research in cancer biology, stem cell differentiation, and high-throughput drug target validation. For further information or to discuss custom applications, please contact Ascent Research.
