The ARAF Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HAP1 cell line, with targeted disruption of the ARAF gene. This heterogeneous pool provides a loss-of-function model that captures the diversity of editing outcomes, making it ideal for studies where clonal variation must be avoided.
HAP1 is a near-haploid chronic myeloid leukemia cell line established from the KBM-7 parental line, originating from a male patient. Its near-haploid genome simplifies genetic manipulation and facilitates clean knockout generation, while its leukemic background maintains relevance for cancer research, particularly hematological malignancies.
ARAF encodes a serine/threonine kinase that functions as a proto-oncogene in the MAPK/ERK signaling cascade. Upon activation by RAS GTPases (HRAS, KRAS, NRAS) following stimulation of receptor tyrosine kinases, ARAF is recruited to the plasma membrane, where it binds 14-3-3 proteins and forms heterodimers with BRAF or CRAF. This activation enables ARAF to phosphorylate the dual-specificity kinases MEK1/2, which subsequently phosphorylate ERK1/2. Activated ERK translocates to the nucleus to regulate transcription of genes promoting cell proliferation and survival. ARAF stability is modulated by HSP90, while the negative regulator PEBP1/RKIP can suppress pathway activity. Additionally, SRC-mediated phosphorylation integrates signals from multiple receptor systems, fine-tuning ARAF activity.
In HAP1 cells, ARAF knockout disrupts MAPK/ERK signaling, potentially blunting oncogenic growth signals inherent to this leukemia model. This allows researchers to dissect ARAF-specific contributions to leukemogenesis and explore pathway crosstalk, such as with PI3K-AKT, in the absence of functional ARAF. The polyclonal nature helps reveal consistent pathway dependencies without clonal bias.
These cells support diverse applications including kinase inhibitor profiling (e.g., MEK or RAF inhibitor dose-response assays), functional genomics screens, and signal transduction studies. Researchers can validate target engagement via western blotting for phospho-MEK and phospho-ERK, phospho-kinase arrays, proliferation assays, and flow cytometry for apoptosis or cell cycle analysis. For further details, contact Ascent Research.
