GPR180 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population for disruption of the GPR180 gene. This product provides a heterogeneous pool of edited cells, each with random genomic modifications at the targeted locus, supplied ready for functional studies. The polyclonal format captures editing diversity typical of pooled CRISPR screens and is ideal for robust loss-of-function phenotypes without clonal selection bias.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, displaying fibroblast-like morphology. Its stable haploid karyotype in most cells simplifies genetic manipulation and knockout phenotype interpretation by eliminating complications from heterozygous alleles. Widely used in CRISPR functional genomics, HAP1 offers genetic tractability and well-characterized biology for signal transduction and drug discovery research.
GPR180 is an orphan GPCR coupling to G??12/13 proteins. Activated by TGF-??, sphingosine-1-phosphate, or lysophosphatidic acid, it stimulates RhoGEF-mediated RhoA activation, leading to ROCK-dependent phosphorylation of LIMK and cofilin to promote actin polymerization. It also activates the MAPK/ERK cascade, causing ERK1/2 phosphorylation and SRF-mediated transcription of genes like CTGF. Interaction with ??-arrestin modulates signaling termination. Beyond vascular remodeling, GPR180 has been linked to energy homeostasis, connecting it to metabolic syndrome.
Knocking out GPR180 in HAP1 cells eliminates G??12/13?CRhoA?CROCK and MAPK/ERK signaling, disrupting cytoskeletal rearrangements and transcriptional responses. In the haploid background, the absence of wild-type alleles ensures strong phenotype penetrance. This polyclonal population is valuable for dissecting GPCR?CTGF-?? crosstalk, mechanisms of vascular remodeling, fibrosis, and metabolic disorders, and enables target validation in hypertension and atherosclerosis research.
Applications include Western blotting for phospho-ERK, RhoA activation assays, RT-qPCR for CTGF, Transwell migration, EdU proliferation, immunofluorescence for stress fibers, and SRF luciferase reporter assays. These cells support vascular smooth muscle biology, GPCR signaling analysis, and drug target validation. For additional information, please contact Ascent Research.
