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Application: Useful for expressing specific HLA-A, B, or C alleles for antigen presentation.
Background: HLA (Human Leukocyte Antigens)-A, B, and C are the three major types of MHC (major histocompatibility complex) class 1 transmembrane proteins. They form a heterodimer with the β2 microglobulin protein (encoded by the B2M gene). The MHC class 1 molecules present short polypeptides, usually between 7-11 amino acids long, to the immune system for recognition as either “self” or “non-self”. HLA-C, for instance, is present in all cells and exists as several haplotypes due to the diversity of HLA-C genes. C*08:02 represents one such haplotype. HLA class I present neoantigen-derived peptides to the cell surface, allowing them to be recognized by T cells, via TCR (T cell receptors). Cancer immunotherapy has been taking advantage of this mechanism by engineering T cells to express TCRs able to recognize specific cancer immunogens. In 2016 the use of HLA-C*08:02-restricted TIL (tumor infiltrating lymphocytes) specifically targeting KRAS (Kirsten rat sarcoma virus) G12D mutation in lung cancer led to positive results. A similar approach was pursued in a patient with metastatic pancreatic cancer and resulted in regression of the disease. The study of HLA-C*08:02-restricted TIL expressing TCR against other neoantigens may prove beneficial in cancer therapy. HLA mismatching between donor cells and the individual can lead to immune rejection, and one option is the knockout of the endogenous HLA, allowing cells to be universally used. The use of a cell line, in which the three major MHC class 1 proteins are not expressed, can be used a cellular model that can be used to express specific HLA alleles.
Description: The HLA-A/B/C Knockout HEK293T Cell Line is a clonal HEK293T cell line that does not express Human Leukocyte Antigen (HLA)-A, HLA-B, and HLA-C. This knockout cell line was generated by CRISPR/Cas9 genome editing using sgRNAs that target regions common to all three genes. This cell line has been validated by flow cytometry to confirm the knockout, as well as HLA-A, B, and C rescue experiments.
Host Cell Line: HEK293T
Mycoplasma Testing: The cell line has been screened to confirm the absence of Mycoplasma species.
Storage Stability: Cells are shipped in dry ice and should immediately be thawed or stored in liquid nitrogen upon receipt. Do not use a -80°C freezer for long term storage.
Supplied As: Each vial contains ≥1 x 106 cells in 1 ml of Cell Freezing Medium (BPS Bioscience #79796)
Target: HLA
Uniprot: P04439, P01889, P10321
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: Leidner R., et al., 2022 N Engl J Med 386:2112-2119.Tran E., et al., 2016 N Eng J Med 375:2255-2262
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