The HM13 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-mediated gene-disrupted polyclonal population derived from the HAP1 cell line. This product offers a robust loss-of-function system for exploring the biological roles of HM13, utilizing a pool of edited cells that collectively lack functional HM13 expression without single-cell cloning.
HAP1 is a near-haploid human adherent cell line originally isolated from the KBM-7 chronic myeloid leukemia (CML) line. Its haploid karyotype enables efficient single-allele gene disruption, yielding complete knockout phenotypes and minimizing compensatory effects from residual gene copies. The CML origin additionally provides a disease-relevant backdrop for investigating leukemia and transplant immunology.
HM13, also known as signal peptide peptidase (SPP), is an intramembrane aspartyl protease residing in the endoplasmic reticulum (ER) membrane. Following cleavage of signal peptides by signal peptidase, SPP processes the remaining fragments, generating peptides capable of binding to HLA-E. This pathway is influenced by ER stress and cytokines, and SPP interacts with the signal peptidase complex, MHC class I molecules, and ER chaperones such as calnexin and calreticulin. Downstream, HM13 activity modulates HLA-E surface expression and MHC class I peptide loading, directly impacting NK cell recognition and minor histocompatibility antigen presentation.
Within HAP1 cells, HM13 disruption provides a precise model to dissect the role of signal peptide processing in CML and immune surveillance. The generation of minor histocompatibility antigens from SPP-cleaved peptides is critical in allogeneic transplantation settings, influencing graft-versus-host disease and graft-versus-leukemia responses. By ablating HM13 in the near-haploid HAP1 background, researchers can unambiguously link changes in the HLA-E peptidome to SPP loss, facilitating the identification of novel peptide antigens and the study of NK cell?Cmediated cytotoxicity.
This polyclonal knockout model supports diverse applications, including minor histocompatibility antigen discovery, transplantation immunology, and cancer immunotherapy target validation. Experimental readouts may involve western blotting and RT-qPCR for HM13 knockout confirmation, flow cytometric analysis of HLA-E surface expression, mass spectrometry-based peptidomics for peptide profiling, and functional assays measuring T cell activation or NK cell cytotoxicity. For additional details, please contact Ascent Research.
