Enhanced Cytosolic Delivery and Release of CRISPR/Cas9 by Black Phosphorus Nanosheets for Genome Editing

Wenhua Zhou; Haodong Cui; Liming Ying; Xue-Feng Yu

doi:10.1002/anie.201806941

Enhanced Cytosolic Delivery and Release of CRISPR/Cas9 by Black Phosphorus Nanosheets for Genome Editing

Angew Chem Int Ed Engl. 2018 Aug 6;57(32):10268-10272. doi: 10.1002/anie.201806941. Epub 2018 Jul 13.

Authors

Wenhua Zhou¹, Haodong Cui¹, Liming Ying², Xue-Feng Yu¹

Affiliations

¹ Center for Biomedical Materials and Interfaces, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, P. R. China.
² Molecular Medicine, National Heart and Lung Institute, Imperial College London, London, SW7 2AZ, UK.

PMID: 29939484
DOI: 10.1002/anie.201806941

Abstract

A biodegradable two-dimensional (2D) delivery platform based on loading black phosphorus nanosheets (BPs) with Cas9 ribonucleoprotein engineered with three nuclear localization signals (NLSs) at C terminus (Cas9N3) is successfully established. The Cas9N3-BPs enter cells effectively via membrane penetration and endocytosis pathways, followed by a BPs biodegradation-associated endosomal escape and cytosolic releases of the loaded Cas9N3 complexes. The Cas9N3-BPs thus provide efficient genome editing and gene silencing in vitro and in vivo at a relatively low dose as compared with other nanoparticle-based delivery platforms. This biodegradable 2D delivery platform offers a versatile cytosolic delivery approach for CRISPR/Cas9 ribonucleoprotein and other bioactive macromolecules for biomedical applications.

Keywords: CRISPR/Cas9; black phosphorus; gene silencing; genome editing; two-dimensional materials.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics*
Cytosol / chemistry
Cytosol / metabolism*
Gene Editing*
Gene Transfer Techniques*
Nanoparticles / chemistry*
Phosphorus / chemistry*

Substances

Phosphorus