The HIF1AN knockout HAP1 polyclonal cells represent a CRISPR/Cas9-edited polyclonal knockout cell population for loss-of-function analysis of HIF1AN. This heterogeneous pool of HAP1 cells carries targeted gene disruptions introduced by CRISPR/Cas9, without clonal isolation. The polyclonal format enables rapid generation of a functional knockout model suitable for pooled screens and biochemical assays requiring a mixed population.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, displaying an adherent, fibroblast-like morphology. Their near-haploid karyotype allows straightforward genetic manipulation by avoiding biallelic editing, making them an ideal host for CRISPR-based functional genomics and phenotypic screens. This property ensures direct genotype?Cphenotype correlations in knockout studies.
HIF1AN encodes factor-inhibiting HIF1A (FIH1), an asparaginyl hydroxylase that represses HIF1A transcriptional activity under normoxia by hydroxylating Asn803, thereby blocking p300/CBP coactivator recruitment. During hypoxia, reduced hydroxylation permits HIF1A to heterodimerize with HIF1B (ARNT) and induce target genes such as VEGF, GLUT1, and EPO, driving angiogenesis, glycolysis, and erythropoiesis. HIF1AN also regulates ankyrin repeat domain proteins like Notch and I??B??. Upstream inputs include oxygen, Fe2+, and 2-oxoglutarate, while interacting factors encompass HIF1A, p300/CBP, and VHL. This positions HIF1AN at a critical junction of oxygen sensing and transcriptional control.
In HAP1 cells, HIF1AN disruption leads to constitutive HIF1A activation, providing a clean model to study hypoxia signaling without external hypoxic stimuli. The near-haploid background simplifies interpretation of HIF1A-dependent phenotypes, allowing investigation of downstream processes such as cell migration, invasion, and metabolic shifts.
Applications include hypoxia signaling research, cancer angiogenesis, metabolic adaptation, and drug target validation. Compatible techniques are Western blot for HIF1A, RT-qPCR for VEGF/GLUT1, HRE-luciferase reporter, ChIP-qPCR, immunofluorescence, hypoxia exposure, migration assays, and co-immunoprecipitation of HIF1A?CHIF1AN complexes. For more information, contact Ascent Research.
