The CD6 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the human near-haploid HAP1 cell line. Engineered to disrupt the CD6 gene, which encodes the T-cell costimulatory receptor CD6, this heterogeneous pool provides a loss-of-function model without clonal selection. The near-haploid genomic architecture is maintained, allowing straightforward functional genomic studies. Researchers can employ these cells to dissect CD6-mediated signaling in a simplified genetic background.
HAP1 cells originate from the KBM-7 chronic myeloid leukemia (CML) line and possess a near-haploid karyotype, facilitating insertional mutagenesis screens, drug-gene profiling, and CRISPR knockout studies. Unlike diploid models, only one allele must be disrupted for functional knockout, enhancing screening efficiency. The parental line expresses a basic hematopoietic program and lacks complex immune signaling, making it ideal for clean pathway interrogation. High transfectability and stable growth further support high-throughput applications.
CD6 is a type I transmembrane glycoprotein of the scavenger receptor cysteine-rich superfamily, primarily expressed on T cells. Upon binding CD166/ALCAM, CD6 co-stimulates TCR signaling by associating with the TCR/CD3 complex and recruiting kinases Lck, Fyn, and ZAP70, along with adaptor GRB2. This triggers ERK phosphorylation and PI3K/AKT activation, promoting T-cell proliferation, survival, and IL-17 production. CD6 expression is regulated by TCR stimulation, IL-2, and Wnt/??-catenin?CTCF-1/LEF-1 inputs. Aberrant CD6 activity enhances Th17 differentiation and contributes to autoimmune disorders such as multiple sclerosis and psoriasis.
In the HAP1 context, CD6 disruption enables dissection of costimulatory signals without confounding T-cell?Cintrinsic networks. Ectopic CD6 reconstitution or downstream reporter assays allow deconvolution of ERK/AKT activation modules in a clean genetic background. The near-haploid state enables genome-wide CRISPR modifier screens to identify synthetic lethal partners, suppressors, or novel CD6 interactors. Thus, this knockout model supports both targeted pathway analysis and unbiased discovery, advancing research into CD6 biology and therapeutic targeting in autoimmunity and lymphoproliferative diseases.
These cells support a variety of experimental applications. Gene disruption can be validated by sequencing or T7E1 mismatch detection, complemented by immunoblotting and flow cytometry for protein loss. Functional readouts include adhesion to CD166, phospho-ERK/AKT analysis, and RT-qPCR of targets like IL17A. Use cases span T-cell costimulation studies, autoimmune disease modeling, drug target validation, haploid screens, and ligand discovery. For further information, please contact Ascent Research.
