The ATRN Knockout HAP1 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal cell population with targeted disruption of the ATRN gene locus. This knockout model is generated in the HAP1 cell background and is provided as a heterogeneous pool of edited cells, enabling direct interrogation of attractin function without the confounding effects of clonal variability. The polyclonal format preserves locus-specific genetic diversity while uniformly abolishing ATRN expression, making it suitable for pooled functional screens and bulk biochemical analyses.
HAP1 cells are a near-haploid, adherent cell line derived from the KBM-7 chronic myeloid leukemia isolate. Their largely haploid genome simplifies loss-of-function studies by eliminating issues of allele compensation, ensuring that CRISPR-mediated disruption of a single allele results in a clear null phenotype. Originating from a male donor, HAP1 cells maintain key signaling networks relevant to hematopoietic and immune contexts, providing a physiologically pertinent platform for dissecting gene function in pathways associated with myeloproliferation and immunological disorders.
ATRN encodes attractin, an accessory receptor for melanocortin receptors (MC1R, MC3R, MC4R) that fine-tunes signal transduction downstream of ligands such as ??-melanocyte-stimulating hormone (??-MSH), agouti-related protein (AGRP), and agouti signaling protein (ASIP). Upon ligand binding, attractin modulates the activation of adenylyl cyclase, leading to cAMP production and subsequent PKA/CREB pathway engagement, as well as ERK/MAPK cascade activity. In immune cells, attractin interacts with the T cell receptor (TCR) complex and CD4, facilitating T cell activation by coupling to downstream kinases including ZAP70 and LAT, ultimately promoting NFAT-mediated transcription and IL-2 cytokine production. This positions ATRN at the intersection of melanocortin signaling, immune cell adhesion, and T cell receptor signaling pathways.
In the HAP1 background, loss of attractin function offers a unique model to study melanocortin-related pathways in a simplified genomic context. The near-haploid nature reduces genetic buffering, making phenotypic outcomes more pronounced and easily quantifiable. Moreover, because HAP1 cells retain functional TCR downstream signaling components, this knockout population can be employed to dissect the role of attractin in coupling melanocortin signal modulation to immune cell activation events, bridging cutaneous biology and hematologic research.
The ATRN Knockout HAP1 Polyclonal Cells are well suited for a range of experimental applications. Functional studies of melanocortin signaling can be performed using cAMP measurement assays, while the impact on immune cell adhesion and T cell activation may be assessed via flow cytometry, cytokine secretion profiling (e.g., IL-2 ELISA), and cell adhesion assays. The polyclonal pool facilitates genetic screens for modifiers of attractin function, and gene disruption can be confirmed by Western blotting and RT-qPCR. This reagent is an invaluable tool for researchers investigating hyperpigmentation, obesity, and immune dysfunction, enabling mechanistic dissection of attractin??s dual roles. For additional technical details or to place an order, please contact Ascent Research.
