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@ARTICLE{Raghavan:296127,
author = {R. Raghavan and M. J. Friedrich and I. King and S.
Chau-Duy-Tam Vo and D. Strebinger and B. Lash and M.
Kilian$^*$ and M. Platten$^*$ and R. K. Macrae and Y. Song
and L. Nivon and F. Zhang},
title = {{R}ational engineering of minimally immunogenic nucleases
for gene therapy.},
journal = {Nature Communications},
volume = {16},
number = {1},
issn = {2041-1723},
address = {[London]},
publisher = {Nature Publishing Group UK},
reportid = {DKFZ-2025-00057},
pages = {105},
year = {2025},
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.},
keywords = {Gene Editing: methods / Humans / CRISPR-Cas Systems /
Genetic Therapy: methods / Animals / CRISPR-Associated
Protein 9: metabolism / CRISPR-Associated Protein 9:
genetics / Protein Engineering: methods / Proprotein
Convertase 9: immunology / Proprotein Convertase 9: genetics
/ Proprotein Convertase 9: metabolism / Mice / HEK293 Cells
/ T-Lymphocytes, Cytotoxic: immunology / Epitopes:
immunology / Endonucleases: metabolism / Endonucleases:
genetics / Female / CRISPR-Associated Protein 9 (NLM
Chemicals) / Proprotein Convertase 9 (NLM Chemicals) /
Epitopes (NLM Chemicals) / Endonucleases (NLM Chemicals) /
PCSK9 protein, human (NLM Chemicals)},
cin = {D170},
ddc = {500},
cid = {I:(DE-He78)D170-20160331},
pnm = {314 - Immunologie und Krebs (POF4-314)},
pid = {G:(DE-HGF)POF4-314},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:39747875},
pmc = {pmc:PMC11696374},
doi = {10.1038/s41467-024-55522-1},
url = {https://inrepo02.dkfz.de/record/296127},
}