This CRISPR/Cas9-edited polyclonal knockout cell population targets CD109 in near-haploid human HAP1 cells. The product comprises a heterogeneous pool of cells bearing disruptive edits, enabling loss-of-function studies without single-cell cloning. CD109 encodes a GPI-anchored glycoprotein that serves as a TGF-?? co-receptor and negative regulator of canonical TGF-?? signaling. The polyclonal format is ideal for functional genomics screens, pathway interrogation, and validation experiments focused on population-level effects of CD109 disruption.
HAP1 cells are a near-haploid human line derived from the KBM-7 chronic myeloid leukemia cell line. Their haploid karyotype facilitates efficient CRISPR/Cas9-mediated gene disruption and avoids diploid gene redundancy. This genetic simplicity has established HAP1 as a widely used model for high-throughput functional screens, systematic knockout studies, and mechanistic dissection of signal transduction, cancer biology, and drug targets. The cell line retains core signaling pathways relevant to both leukemia and solid tumors.
CD109 is a cell-surface GPI-anchored protein that modulates TGF-?? signaling by acting as an accessory receptor. It binds TGF-?? ligands (TGFB1, TGFB2, TGFB3) and associates with TGF-?? receptor type I (TGFBR1) and type II (TGFBR2) to inhibit phosphorylation of SMAD2 and SMAD3. This negative regulation attenuates SMAD-dependent transcription, reducing expression of target genes such as SERPINE1 (PAI-1), COL1A1, and FN1. CD109 also interacts with the co-receptor TGFBR3 and may influence non-canonical JAK-STAT and MAPK pathways, thereby regulating cell proliferation, migration, and immune responses.
In HAP1 cells, CD109 knockout provides a clean genetic background to dissect its regulatory role in TGF-??-driven processes. The near-haploid nature ensures a robust loss-of-function phenotype in the polyclonal population without clonal bias. This model is especially relevant for studying CD109 in cancers where its expression is altered, including squamous cell carcinoma, glioblastoma, melanoma, and lung cancer. Researchers can directly assess effects on SMAD2/3 phosphorylation kinetics, TGF-??-induced transcriptional programs, and downstream cellular behaviors like proliferation and migration.
This polyclonal knockout product supports quantitative TGF-?? pathway analysis via luciferase reporter assays, phospho-SMAD2/3 Western blotting, and RT-qPCR for target genes (e.g., SERPINE1). Functional assays for proliferation and migration link CD109 loss to altered tumorigenic properties. The model also enables flow cytometry, co-culture experiments, and high-throughput screens for TGF-?? signaling modulators. For further technical details and custom options, contact Ascent Research.
