Rational engineering of minimally immunogenic nucleases for gene therapy
Abstract Genome editing using CRISPR-Cas systems is a promising avenue for the treatment of genetic diseases. However, cellular and humoral immunogenicity of genome editing tools, which originate from bacteria, complicates their clinical use. Here we report reduced immunogenicity (Red)(i)-variants o...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55522-1 |
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| author | Rumya Raghavan Mirco J. Friedrich Indigo King Samuel Chau-Duy-Tam Vo Daniel Strebinger Blake Lash Michael Kilian Michael Platten Rhiannon K. Macrae Yifan Song Lucas Nivon Feng Zhang |
| author_facet | Rumya Raghavan Mirco J. Friedrich Indigo King Samuel Chau-Duy-Tam Vo Daniel Strebinger Blake Lash Michael Kilian Michael Platten Rhiannon K. Macrae Yifan Song Lucas Nivon Feng Zhang |
| author_sort | Rumya Raghavan |
| collection | DOAJ |
| description | Abstract Genome editing using CRISPR-Cas systems is a promising avenue for the treatment of genetic diseases. However, cellular and humoral immunogenicity of genome editing tools, which originate from bacteria, complicates their clinical use. Here we report reduced immunogenicity (Red)(i)-variants of two clinically relevant nucleases, SaCas9 and AsCas12a. Through MHC-associated peptide proteomics (MAPPs) analysis, we identify putative immunogenic epitopes on each nuclease. Using computational modeling, we rationally design these proteins to evade the immune response. SaCas9 and AsCas12a Redi variants are substantially less recognized by adaptive immune components, including reduced binding affinity to MHC molecules and attenuated generation of cytotoxic T cell responses, yet maintain wild-type levels of activity and specificity. In vivo editing of PCSK9 with SaCas9.Redi.1 is comparable in efficiency to wild-type SaCas9, but significantly reduces undesired immune responses. This demonstrates the utility of this approach in engineering proteins to evade immune detection. |
| format | Article |
| id | doaj-art-5d8514c553eb4cc78b4307dec796613f |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-5d8514c553eb4cc78b4307dec796613f2025-01-05T12:38:40ZengNature PortfolioNature Communications2041-17232025-01-0116111310.1038/s41467-024-55522-1Rational engineering of minimally immunogenic nucleases for gene therapyRumya Raghavan0Mirco J. Friedrich1Indigo King2Samuel Chau-Duy-Tam Vo3Daniel Strebinger4Blake Lash5Michael Kilian6Michael Platten7Rhiannon K. Macrae8Yifan Song9Lucas Nivon10Feng Zhang11Broad Institute of MIT and HarvardBroad Institute of MIT and HarvardCyrus BiotechnologyBroad Institute of MIT and HarvardBroad Institute of MIT and HarvardBroad Institute of MIT and HarvardClinical Cooperation Unit Neuroimmunology and Brain Tumor Immunology, German Cancer Research Center, DKFZClinical Cooperation Unit Neuroimmunology and Brain Tumor Immunology, German Cancer Research Center, DKFZBroad Institute of MIT and HarvardCyrus BiotechnologyCyrus BiotechnologyBroad Institute of MIT and HarvardAbstract Genome editing using CRISPR-Cas systems is a promising avenue for the treatment of genetic diseases. However, cellular and humoral immunogenicity of genome editing tools, which originate from bacteria, complicates their clinical use. Here we report reduced immunogenicity (Red)(i)-variants of two clinically relevant nucleases, SaCas9 and AsCas12a. Through MHC-associated peptide proteomics (MAPPs) analysis, we identify putative immunogenic epitopes on each nuclease. Using computational modeling, we rationally design these proteins to evade the immune response. SaCas9 and AsCas12a Redi variants are substantially less recognized by adaptive immune components, including reduced binding affinity to MHC molecules and attenuated generation of cytotoxic T cell responses, yet maintain wild-type levels of activity and specificity. In vivo editing of PCSK9 with SaCas9.Redi.1 is comparable in efficiency to wild-type SaCas9, but significantly reduces undesired immune responses. This demonstrates the utility of this approach in engineering proteins to evade immune detection.https://doi.org/10.1038/s41467-024-55522-1 |
| spellingShingle | Rumya Raghavan Mirco J. Friedrich Indigo King Samuel Chau-Duy-Tam Vo Daniel Strebinger Blake Lash Michael Kilian Michael Platten Rhiannon K. Macrae Yifan Song Lucas Nivon Feng Zhang Rational engineering of minimally immunogenic nucleases for gene therapy Nature Communications |
| title | Rational engineering of minimally immunogenic nucleases for gene therapy |
| title_full | Rational engineering of minimally immunogenic nucleases for gene therapy |
| title_fullStr | Rational engineering of minimally immunogenic nucleases for gene therapy |
| title_full_unstemmed | Rational engineering of minimally immunogenic nucleases for gene therapy |
| title_short | Rational engineering of minimally immunogenic nucleases for gene therapy |
| title_sort | rational engineering of minimally immunogenic nucleases for gene therapy |
| url | https://doi.org/10.1038/s41467-024-55522-1 |
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