Multifunctional CRISPR-Cas9 with engineered immunosilenced human T cell epitopes

• DOI: 10.1038/s41467-019-09693-x
• Publication Date: 2019-04-23T10:04:03Z
• Authors: Shayesteh R. Ferdosi; Radwa Ewaisha; Farzaneh Moghadam; Sri Krishna; Jin G. Park; Mo R. Ebrahimkhani; Samira Kiani; Karen S. Anderson
• Keywords: HLA-A Antigens; Streptococcus pyogenes; Science; Q; Antigen-Presenting Cells; Epitopes, T-Lymphocyte; Genetic Therapy; CD8-Positive T-Lymphocytes; Lymphocyte Activation; Antibodies, Bacterial; Article; Healthy Volunteers; 3. Good health; HEK293 Cells; Mutagenesis; CRISPR-Associated Protein 9; Humans; Precision Medicine; Cell Engineering; Epitope Mapping

The CRISPR-Cas9 system has raised hopes for developing personalized gene therapies complex diseases. Its application genetic and epigenetic in humans raises concerns over immunogenicity of the bacterially derived Cas9 protein. Here we detect antibodies to Streptococcus pyogenes (SpCas9) at least 5% 143 healthy individuals. We also report pre-existing human CD8+T cell immunity majority individuals screened. identify two immunodominant SpCas9 T epitopes HLA-A*02:01 using an enhanced prediction algorithm that incorporates receptor contact residue hydrophobicity HLA binding evaluated them by assays donor PBMCs. In a proof-of-principle study, demonstrate protein can be modified eliminate through targeted mutation while preserving its function specificity. Our study highlights problem against CRISPR-associated nucleases offers potential solution mitigate immune response.