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Investigation of Cas9 antibodies in the human eye
Journal article   Open access   Peer reviewed

Investigation of Cas9 antibodies in the human eye

Marcus A Toral, Carsten T Charlesworth, Benjamin Ng, Teja Chemudupati, Shota Homma, Hiromitsu Nakauchi, Alexander G Bassuk, Matthew H Porteus and Vinit B Mahajan
Nature communications, Vol.13(1), pp.1053-1053
02/25/2022
DOI: 10.1038/s41467-022-28674-1
PMCID: PMC8881612
PMID: 35217666
url
https://doi.org/10.1038/s41467-022-28674-1View
Published (Version of record) Open Access

Abstract

Preexisting immunity against Cas9 proteins in humans represents a safety risk for CRISPR-Cas9 technologies. However, it is unclear to what extent preexisting Cas9 immunity is relevant to the eye as it is targeted for early in vivo CRISPR-Cas9 clinical trials. While the eye lacks T-cells, it contains antibodies, cytokines, and resident immune cells. Although precise mechanisms are unclear, intraocular inflammation remains a major cause of vision loss. Here, we used immunoglobulin isotyping and ELISA platforms to profile antibodies in serum and vitreous fluid biopsies from human adult subjects and Cas9-immunized mice. We observed high prevalence of preexisting Cas9-reactive antibodies in serum but not in the eye. However, we detected intraocular antibodies reactive to S. pyogenes-derived Cas9 after S. pyogenes intraocular infection. Our data suggest that serum antibody concentration may determine whether specific intraocular antibodies develop, but preexisting immunity to Cas9 may represent a lower risk in human eyes than systemically.

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