The HDGF Knockout HAP1 Polyclonal Cells consist of a CRISPR/Cas9-edited polyclonal knockout cell population targeting the human HDGF (hepatoma-derived growth factor) gene in the HAP1 host cell line. This loss-of-function model is designed for studying HDGF-dependent cellular processes, enabling investigation of its roles in proliferation, survival, and transcriptional regulation. The polyclonal format provides a mixed population of edited alleles, suitable for pooled loss-of-function screens and pooled functional analyses.
HAP1 is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia (CML) line. Its haploid karyotype makes it an ideal platform for genetic screening and CRISPR/Cas9-based functional genomics, as it simplifies the generation and interpretation of knockout phenotypes. The cells maintain key signaling pathways relevant to CML and solid tumor biology, providing a physiologically relevant context for cancer research.
HDGF encodes a heparin-binding growth factor with mitogenic and angiogenic properties, also functioning as a transcriptional repressor of cell cycle inhibitors such as p21. It is activated by upstream signals including EGF, PDGF, TGF-??, and hypoxia via HIF-1??, and regulated by transcription factors E2F and NF-??B. Upon stimulation, HDGF interacts with nucleolin and importin ?? and requires heparin for nuclear translocation. Mechanistically, HDGF promotes cell proliferation and survival by activating PI3K/AKT and MAPK/ERK cascades, leading to upregulation of cyclin D1, c-Myc, VEGF, Bcl-2, and survivin. In the nucleus, it represses p21 transcription, thereby overriding cell cycle checkpoints.
In the HAP1 background, the HDGF knockout population enables dissection of these signaling modules in a genetically tractable system. Given HDGF’s overexpression in hepatocellular carcinoma, non-small cell lung cancer, gastric cancer, colorectal cancer, and glioma, this model supports studies of oncogenic signaling, growth factor dependence, and apoptotic resistance. It is particularly useful for examining cross-talk between HDGF-controlled pathways (such as EGFR??RAS??RAF??MEK??ERK and PI3K/AKT/mTOR) and the Wnt/??-catenin/TCF/LEF axis, as well as for screening compounds that target HDGF-mediated survival signals.
Researchers can employ this knockout polyclonal population in a range of assays including MTS/MTT proliferation assays, western blotting for phosphorylated ERK and AKT, RT-qPCR quantification of downstream targets (cyclin D1, c-Myc, VEGF), and transwell migration/invasion experiments. Additional applications include ChIP-qPCR to assess HDGF DNA binding, immunofluorescence localization studies, flow cytometric cell cycle analysis, and Annexin V apoptosis assays. These cells are well-suited for drug target validation, angiogenesis studies, and functional genomics screens in cancer biology. For technical support, please contact Ascent Research.
