This CRISPR/Cas9-edited polyclonal knockout cell population targets the human GNRH1 gene in the HAP1 cell line, generating a heterogeneous pool of cells with disrupted gonadotropin-releasing hormone (GnRH) production. The polyclonal format supports pooled genetic screens and bulk biochemical assays without requiring clonal isolation, providing a robust loss-of-function model for GnRH biology.
The HAP1 host is a near-haploid human myeloid cell line derived from the KBM-7 chronic myeloid leukemia line. Its haploid karyotype simplifies gene disruption as most loci are single-copy, enabling efficient CRISPR/Cas9 editing. This genetic tractability makes HAP1 a versatile platform for knockout studies, even for pathways not natively active, through complementation with exogenous constructs.
GNRH1 encodes the precursor of GnRH, a key hypothalamic decapeptide that stimulates pituitary gonadotropin secretion. GnRH binds GNRHR, a G??q-coupled receptor, triggering GNAQ, PLCB, and calcium signaling, which activates PRKCA and MAPK1/ERK2 to promote LHB and FSHB expression. Upstream regulators include KISS1/KISS1R and TAC3/TACR3, with feedback from sex steroids and leptin. Knockout of GNRH1 eliminates GnRH, disrupting the HPG axis and gonadotropin output.
In HAP1 cells, this knockout provides a null background for reconstituting GnRH signaling. Although non-neuronal, HAP1 can express GNRHR and pituitary transcription factors, enabling dissection of receptor activation and downstream gene regulation. The polyclonal pool minimizes background signal and is well-suited for high-throughput CRISPR modifier screens to identify novel pathway components.
Applications encompass reproductive biology, neuroendocrinology, and infertility research. The model supports GnRH ELISA, qRT-PCR for LHB/FSHB, GNRHR calcium flux assays, and reporter gene assays. It is also valuable for synthetic rescue experiments and focused genetic screens. For further information, contact Ascent Research.
