GPR75 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the near-haploid HAP1 cell line, targeting the GPR75 gene locus. This product provides a loss-of-function model for investigating the role of GPR75 in metabolic and inflammatory signaling pathways.
The host cell line, HAP1, is a near-haploid human cell line originally derived from the KBM-7 chronic myeloid leukemia line. With a haploid karyotype, HAP1 cells contain a single copy of most chromosomes, which simplifies genetic manipulation and reduces the likelihood of functional redundancy. These cells exhibit adherent growth and are of male origin, making them a robust and widely used platform for CRISPR-based knockout screens and functional genomics studies.
GPR75 encodes a G protein-coupled receptor that has been implicated in chemokine signaling, particularly as a receptor for CCL5 (RANTES) and potentially chemerin. Mechanistically, GPR75 couples to G??i/o and G??q heterotrimeric G proteins. G??i/o activation inhibits adenylyl cyclase, lowering intracellular cAMP levels, while G??q stimulates phospholipase C?? (PLC??), leading to inositol trisphosphate (IP3)-mediated calcium release and diacylglycerol (DAG)-dependent activation of protein kinase C (PKC). Downstream, these pathways converge on the mitogen-activated protein kinase (MAPK/ERK) cascade and transcription factors such as cAMP response element-binding protein (CREB) and nuclear factor kappa B (NF-??B). Additionally, GPR75 signaling is regulated by G protein-coupled receptor kinases (GRKs) and ??-arrestin, which mediate receptor desensitization and internalization. This signaling network positions GPR75 as a key regulator of energy homeostasis, appetite, and inflammatory responses.
In the near-haploid HAP1 background, disruption of the single GPR75 allele results in a complete loss of protein function, creating an unambiguous loss-of-function phenotype. This model is particularly useful for dissecting GPR75??s role in chemokine-mediated signaling and its impact on metabolic disorders, such as obesity and type 2 diabetes, as well as inflammatory diseases. The polyclonal nature of the knockout population preserves cellular heterogeneity while ensuring the absence of wild-type receptor, thus enabling robust phenotypic screening.
These knockout cells are suited for a wide range of experimental applications. Researchers can employ them in functional assays like cAMP and calcium flux measurements to assess G protein coupling, ERK phosphorylation analysis by flow cytometry to monitor MAPK pathway activation, and CCL5-mediated migration assays to study chemotactic responses. They are also ideal for high-throughput screening of receptor ligands, drug target validation, and transcriptomic or proteomic profiling via RNA-seq. The model supports investigations into the interplay between metabolic and immune signaling pathways, offering a valuable tool for preclinical research. For detailed technical specifications and ordering information, please contact Ascent Research.
