TY  - JOUR
AU  - Raghavan, Rumya
AU  - Friedrich, Mirco J
AU  - King, Indigo
AU  - Chau-Duy-Tam Vo, Samuel
AU  - Strebinger, Daniel
AU  - Lash, Blake
AU  - Kilian, Michael
AU  - Platten, Michael
AU  - Macrae, Rhiannon K
AU  - Song, Yifan
AU  - Nivon, Lucas
AU  - Zhang, Feng
TI  - Rational engineering of minimally immunogenic nucleases for gene therapy.
JO  - Nature Communications
VL  - 16
IS  - 1
SN  - 2041-1723
CY  - [London]
PB  - Nature Publishing Group UK
M1  - DKFZ-2025-00057
SP  - 105
PY  - 2025
AB  - 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.
KW  - Gene Editing: methods
KW  - Humans
KW  - CRISPR-Cas Systems
KW  - Genetic Therapy: methods
KW  - Animals
KW  - CRISPR-Associated Protein 9: metabolism
KW  - CRISPR-Associated Protein 9: genetics
KW  - Protein Engineering: methods
KW  - Proprotein Convertase 9: immunology
KW  - Proprotein Convertase 9: genetics
KW  - Proprotein Convertase 9: metabolism
KW  - Mice
KW  - HEK293 Cells
KW  - T-Lymphocytes, Cytotoxic: immunology
KW  - Epitopes: immunology
KW  - Endonucleases: metabolism
KW  - Endonucleases: genetics
KW  - Female
KW  - CRISPR-Associated Protein 9 (NLM Chemicals)
KW  - Proprotein Convertase 9 (NLM Chemicals)
KW  - Epitopes (NLM Chemicals)
KW  - Endonucleases (NLM Chemicals)
KW  - PCSK9 protein, human (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:39747875
C2  - pmc:PMC11696374
DO  - DOI:10.1038/s41467-024-55522-1
UR  - https://inrepo02.dkfz.de/record/296127
ER  -