biostudies-literatureChen GNEI NIH HHSNHLBI NIH HHSNINDS NIH HHSNIGMS NIH HHS974-980https://www.ebi.ac.uk/biostudies/studies/S-EPMC6778035biostudies-literatureUnknown14(10)Delivery technologies for the CRISPR-Cas9 (CRISPR, clustered regularly interspaced short palindromic repeats) gene editing system often require viral vectors, which pose safety concerns for therapeutic genome editing¹. Alternatively, cationic liposomal components or polymers can be used to encapsulate multiple CRISPR components into large particles (typically >100 nm diameter); however, such systems are limited by variability in the loading of the cargo. Here, we report the design of customizable synthetic nanoparticles for the delivery of Cas9 nuclease and a single-guide RNA (sgRNA) that enables the controlled stoichiometry of CRISPR components and limits the possible safety concerns in vivo. We describe the synthesis of a thin glutathione (GSH)-cleavable covalently crosslinkedNature nanotechnologyA biodegradable nanocapsule delivers a Cas9 ribonucleoprotein complex for in vivo genome editing.PMC6778035R01 HL129785R01 NS091540R01 EY024995R35 GM119644UG3 NS111688R01 HL143469Robertson SGong SSuzuki MShahi PKChen GAbdeen AAPattnaik BRSaha KWang YXie RfalseA biodegradable nanocapsule delivers a Cas9 ribonucleoprotein complex for in vivo genome editing.Delivery technologies for the CRISPR-Cas9 (CRISPR, clustered regularly interspaced short palindromic repeats) gene editing system often require viral vectors, which pose safety concerns for therapeutic genome editing¹. Alternatively, cationic liposomal components or polymers can be used to encapsulate multiple CRISPR components into large particles (typically >100 nm diameter); however, such systems are limited by variability in the loading of the cargo. Here, we report the design of customizable synthetic nanoparticles for the delivery of Cas9 nuclease and a single-guide RNA (sgRNA) that enables the controlled stoichiometry of CRISPR components and limits the possible safety concerns in vivo. We describe the synthesis of a thin glutathione (GSH)-cleavable covalently crosslinked2019-01-01T00:00:00Z2019 Oct2025-04-04T00:05:20.669Z2020-05-22T12:16:01ZS-EPMC67780353150153210.1038/s41565-019-0539-2