HLA-A/B/C Knockout Electroporation Kit

The HLA-A/B/C Knockout Electroporation Kit is suitable for cell line and primary T cell engineering via electroporation. The kit contains both the Cas9 enzyme (Streptococcus pyogenes) and the gRNA targeting HLA-A/B/C (Human Leukocyte Antigens). This kit is sufficient to engineer up to 5 million primary T cells. 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 beta2 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 ?oself?? or ?onon-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 that 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 resulted in 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. K562 cells are HLA class I and II negative, making them an ideal cellular model to introduce and study specific haplotype responses. HLA mismatching between donor cells and the individual can lead to immune rejection, and one option is the knockout the endogenous HLA, allowing cells to be more widely universally used.