ADISSP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HAP1 cell line, with targeted disruption of the ADISSP gene. This loss-of-function model enables investigation of the functional roles of ADISSP, an adipose-secreted protein implicated in regulating insulin sensitivity and metabolic homeostasis. The polyclonal format provides a heterogeneous pool of edited cells, suitable for robust functional assays without clonal isolation, thereby supporting pooled screening and mechanistic studies of ADISSP-dependent signaling.
HAP1 cells are a near-haploid human cell line derived from a male chronic myeloid leukemia patient; they display an adherent, fibroblast-like morphology. The haploid karyotype simplifies genetic manipulation by reducing redundancy, enabling efficient gene disruption and phenotypic analysis. Widely employed in CRISPR-based knockout screens, HAP1 cells retain intact core signaling modules, making them a versatile host for investigating diverse biological pathways. Their genetic stability and ease of culture further recommend them for generating knockout models in genes that may not be endogenously expressed in typical cell lines.
ADISSP is an adipose-secreted factor whose expression is transcriptionally regulated by PPARG, CEBPA, insulin, and SREBF1. Secreted ADISSP is believed to interact with other adipokines such as ADIPOQ, LEP, and RETN, potentially forming regulatory complexes that modulate systemic metabolism. Downstream, ADISSP signaling converges on insulin receptor substrate 1 (IRS1) and the kinase AKT, ultimately affecting translocation of the glucose transporter SLC2A4 (GLUT4) and cellular glucose uptake. Moreover, ADISSP may modulate expression of adiponectin (ADIPOQ), thereby linking its activity to broader adipokine and inflammatory signaling networks.
In the HAP1 context, ADISSP knockout offers a unique platform to dissect crosstalk between adipokines and insulin signaling components. Although HAP1 cells do not naturally secrete adipokines, they can be engineered to express pathway components or used as sensor models upon exogenous stimulation. The haploid background accelerates knockout generation, while the polyclonal population permits assessment of average gene disruption effects without clonal bias. This system enables detailed examination of AKT and IRS1 phosphorylation dynamics and changes in expression of ADIPOQ or SLC2A4 in response to insulin or other metabolic stimuli.
This cell pool is well suited for adipokine signaling studies, insulin sensitivity assays, and metabolic pathway analysis. Representative techniques include Western blotting for protein phosphorylation (e.g., AKT, IRS1), RT-qPCR for ADISSP and target gene expression, glucose uptake assays, and co-immunoprecipitation to detect adipokine interactions. These cells can also be employed in drug screening for metabolic disorders or metabolic profiling to map signaling consequences of ADISSP loss. For additional information and support, please contact Ascent Research.
