The GPR62 Knockout HAP1 Polyclonal Cells are a pooled population of HAP1 cells edited by CRISPR/Cas9 to disrupt the GPR62 gene, which encodes an orphan G protein?coupled receptor. This polyclonal knockout model provides a heterogeneous loss?of?function resource that avoids the clonal artifacts sometimes associated with single?cell?derived lines. The CRISPR/Cas9?mediated gene disruption eliminates wild?type GPR62 expression, generating a functional knockout suitable for a range of biochemical, cell?based, and genomic analyses.
HAP1 is a near?haploid human fibroblast?like cell line derived from the KBM?7 chronic myeloid leukemia lineage. It possesses a haploid karyotype for all chromosomes except chromosome 8, which greatly simplifies genetic analysis because recessive phenotypes are unmasked by the absence of a second allele. HAP1 cells grow adherently, exhibit robust signaling competence, and are a well?established platform for functional genomics, drug?target discovery, and high?throughput CRISPR screens.
GPR62 functions as an orphan G protein?coupled receptor with no confirmed endogenous ligand. It activates Gs and Gi/o proteins, which in turn regulate adenylyl cyclase, generating the second messenger cAMP. cAMP activates protein kinase A (PKA), which phosphorylates the transcription factor CREB, modulating gene expression. In a parallel pathway, GPR62 recruits ???arrestin to activate the MAPK cascade, culminating in phosphorylation of ERK1/2. Receptor activity may be further modulated by GRK?mediated phosphorylation and interactions with RAMPs. Thus, GPR62 functionally intersects the cAMP/PKA/CREB and MAPK/ERK signaling networks.
In the HAP1 near?haploid background, knockout of GPR62 eliminates its contribution to basal and stimulated signaling, enabling clean dissection of receptor?specific functions. The absence of a second allele ensures that any observed phenotype is directly linked to GPR62 loss, reducing background noise. This model is particularly valuable for deorphanization campaigns, where the knockout serves as a negative control for ligand?dependent GPCR responses. Moreover, the polyclonal nature of the population avoids clonal selection biases, making it suitable for large?scale screens such as genome?wide CRISPR modifier screens or drug sensitivity profiling.
Typical assays with these cells include cAMP accumulation measurements using biosensors, calcium flux assays with fluorescent indicators, and ???arrestin recruitment tests such as Tango or PathHunter. Downstream signaling can be probed by Western blotting for phosphorylated CREB and ERK1/2, and global gene expression changes can be assessed by RNA?seq. These polyclonal knockout cells are pertinent to studying neuropsychiatric disorders like autism and schizophrenia, as well as basic orphan receptor biology and drug target identification. For more information or to place an order, please contact Ascent Research.
