Efficient in vivo neuronal genome editing in the mouse brain using nanocapsules containing CRISPR-Cas9 ribonucleoproteins.
Document Type
Article
Publication Date
2-1-2023
Original Citation
Metzger J,
Wang Y,
Neuman S,
Snow KJ,
Murray SA,
Lutz C,
Bondarenko V,
Felton J,
Gimse K,
Xie R,
Li D,
Zhao Y,
Flowers M,
Simmons H,
Roy S,
Saha K,
Levine J,
Emborg M,
Gong S.
Efficient in vivo neuronal genome editing in the mouse brain using nanocapsules containing CRISPR-Cas9 ribonucleoproteins. Biomaterials. 2022;293:121959.
Keywords
JMG, Animals, Mice, Gene Editing, CRISPR-Cas Systems, Nanocapsules, Ribonucleoproteins, Neurons, Brain
JAX Source
Biomaterials. 2022;293:121959.
ISSN
1878-5905
PMID
36527789
DOI
https://doi.org/10.1016/j.biomaterials.2022.121959
Grant
This work was supported by the NIH Common Fund and National Institutes of Health Office of the Director U54 OD026635 (S.A.M. and C.M.L.), the WIMR optical imaging core (1S10OD025040- 01), NIH UG3-NS111688 (S.R., K.S., J.E.L., M.E.E., S.G.), and NIH UH3- NS111688 (S.R., K.S., J.E.L., M.E.E., S.G.).
Abstract
Genome editing of somatic cells via clustered regularly interspaced short palindromic repeats (CRISPR) offers promise for new therapeutics to treat a variety of genetic disorders, including neurological diseases. However, the dense and complex parenchyma of the brain and the post-mitotic state of neurons make efficient genome editing challenging. In vivo delivery systems for CRISPR-Cas proteins and single guide RNA (sgRNA) include both viral vectors and non-viral strategies, each presenting different advantages and disadvantages for clinical application. We developed non-viral and biodegradable PEGylated nanocapsules (NCs) that deliver preassembled Cas9-sgRNA ribonucleoproteins (RNPs). Here, we show that the RNP NCs led to robust genome editing in neurons following intracerebral injection into the healthy mouse striatum. Genome editing was predominantly observed in medium spiny neurons (>80%), with occasional editing in cholinergic, calretinin, and parvalbumin interneurons. Glial activation was minimal and was localized along the needle tract. Our results demonstrate that the RNP NCs are capable of safe and efficient neuronal genome editing in vivo.