The HM13 Knockout Jurkat Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Jurkat human T lymphocyte cell line. This product provides a loss-of-function model of the HM13 gene (encoding signal peptide peptidase-like 3, SPPL3) for advanced biomedical research. The polyclonal format offers a heterogeneous pool of edited cells, enabling robust functional studies of HM13 disruption in a defined immortalized T cell background without clonal selection artifacts.
Jurkat cells are an immortalized human T lymphocyte line originally derived from the peripheral blood of a patient with acute T cell leukemia. They are characterized by p53 deficiency and PTEN-null status, making them a widely employed model for investigating T cell receptor signaling, apoptosis, and leukemogenesis. These features render Jurkat cells particularly suitable for probing the intersection of immune signaling, antigen presentation, and tumor biology.
HM13 encodes the intramembrane aspartyl protease SPPL3, which resides in the endoplasmic reticulum. SPPL3 cleaves transmembrane substrates, including N-terminal fragments of the MHC class I heavy chain, facilitating the generation of antigenic peptides for presentation. Its activity is regulated by cytokines such as IFNG and TNF, acting through transcription factors NFKB1 and IRF1. SPPL3 functions within the peptide loading complex, interacting with TAP1, TAP2, tapasin, calreticulin, ERp57, and calnexin, to influence HLA-A2 peptide repertoires and surface glycosylation.
In the Jurkat T cell context, disruption of HM13 by CRISPR/Cas9 ablates SPPL3 protease function, perturbing MHC class I antigen processing and presentation. This knockout model allows dissection of minor histocompatibility antigen generation, glycosylation-dependent immune recognition, and the contribution of SPPL3 to immune evasion mechanisms relevant to T cell leukemia and alloreactivity. The p53-null and PTEN-null background further accentuates studies of cancer-related immune escape.
Researchers can employ this knockout model in a variety of experimental settings, including flow cytometric analysis of HLA-A2 surface expression, Western blotting of MHC class I components, RT-qPCR profiling of downstream gene targets, and T cell activation assays using NFAT reporters. Additional applications encompass co-immunoprecipitation of peptide loading complexes, MHC peptide elution and mass spectrometry for immunopeptidome analysis, and functional evaluation of glycosylation pathways. These tools support investigations into graft-versus-host disease, minor histocompatibility antigen biology, and cancer immunotherapies. For further information, please contact Ascent Research.
