The CD2 Knockout HAP1 Polyclonal Cells represent a genetically engineered cell population in which the CD2 locus has been disrupted via CRISPR/Cas9-mediated genome editing. This polyclonal knockout product offers a heterogeneous pool of edited cells, suitable for functional genomics experiments and drug screening campaigns. Utilizing the HAP1 near-haploid cell line, this model minimizes genetic redundancy and facilitates unambiguous genotype-phenotype correlation in costimulatory signaling research.
The HAP1 cell line is a near-haploid derivative of the KBM-7 chronic myeloid leukemia cell line, originally derived from a patient with CML. HAP1 cells lack a full diploid genome, rendering them particularly valuable for haploid genetic screens, including CRISPR knockout and insertional mutagenesis studies. Their myeloid lineage background and robust growth characteristics make them a versatile platform for dissecting signaling pathways relevant to immune cell biology, despite not being of T-cell origin. The near-haploid state simplifies the generation and characterization of gene disruptions, allowing researchers to rapidly assess the functional consequences of CD2 loss.
CD2 encodes a transmembrane glycoprotein that serves as a costimulatory receptor on T lymphocytes and natural killer cells. Engagement by CD58 (LFA-3) on antigen-presenting cells enhances TCR signaling by amplifying proximal tyrosine phosphorylation. CD2 interacts with CD2AP and LCK, linking its tail to the LCK/ZAP70 cascade. This costimulation potentiates PI3K/AKT and MAPK pathways, upregulating IL-2 and IFN-??. TCR signals and transcription factors TCF-1 and GATA3 regulate CD2 expression during T-cell development.
Although HAP1 originates from a CML line, ectopic CD2 expression creates a simplified model for studying CD2 costimulatory function. The near-haploid genome eliminates allelic redundancy, enabling clear assignment of phenotypic effects to CD2 disruption. This system is suited for biochemical dissection of CD2-proximal signaling, such as LCK phosphorylation and formation of LAT/SLP-76/PLC??1 complexes. The polyclonal population better reflects natural immune cell heterogeneity, enhancing relevance for pharmacological studies.
Researchers can employ the CD2 Knockout HAP1 Polyclonal Cells in a variety of experimental settings, including T-cell adhesion assays on CD58-coated surfaces, flow cytometric assessment of CD2 surface expression, Western blotting for CD2 protein and its downstream targets (e.g., phospho-LCK, phospho-ZAP70), and cytokine quantification by ELISA following TCR stimulation. Transcriptomic analysis via RNA-seq can reveal gene expression changes attributable to CD2 loss, while functional screens can identify synthetic lethal interactions or modulators of CD2-mediated costimulation. These applications support drug discovery efforts targeting CD2?CCD58 interactions in autoimmune disorders, transplant rejection, and certain lymphoproliferative diseases. For further technical details and ordering information, please contact Ascent Research.
