The AMBP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population produced in the human HAP1 cell line, designed to create a loss-of-function model for the AMBP gene. This population enables study of the bifunctional AMBP protein without clonal isolation.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line, extensively utilized in haploid genetic screens due to its single allele per gene. The haploid state allows efficient CRISPR/Cas9-mediated gene disruption and simplifies interpretation of knockout phenotypes, making it a powerful host for studying genes involved in stress responses and signaling.
The AMBP gene encodes a precursor that is processed into two proteins: alpha-1-microglobulin (A1M) and bikunin. A1M is a heme-binding radical scavenger with antioxidant properties, protecting cells from oxidative damage and heme toxicity. Bikunin is a Kunitz-type serine protease inhibitor that targets trypsin, plasmin, and neutrophil elastase, modulating inflammation and extracellular matrix stability. AMBP transcription is activated by HNF1A and C/EBP-??, and upregulated by pro-inflammatory cytokines such as IL-6. A1M interacts with heme and albumin, while bikunin forms complexes with inter-alpha-trypsin inhibitor heavy chains, integrating AMBP into pathways of heme detoxification, redox balance, and protease inhibition.
Disruption of AMBP in the near-haploid HAP1 background eliminates both functional subunits, creating a clean genetic model to dissect the individual and combined contributions of A1M and bikunin. This polyclonal knockout population is particularly valuable for investigating cellular responses to heme overload and oxidative stress, as well as for studying the role of bikunin in controlling serine protease activity. Relevant disease models include acute kidney injury, hemoglobinuria, and preeclampsia, where dysregulation of heme scavenging and protease inhibition are central.
Researchers can validate knockout by western blotting for A1M and bikunin or RT-qPCR for AMBP mRNA. Functional assays include heme-binding and reactive oxygen species measurement for A1M, trypsin inhibition for bikunin, and cell viability under oxidative stress. Additional studies may explore downstream inflammatory signaling. This AMBP knockout tool supports research in antioxidant defense, heme biology, and inflammatory disorders. For technical inquiries, please contact Ascent Research.
