Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein for Efficient Gene Editing

Rubul Mout; Moumita Ray; Gulen Yesilbag Tonga; Yi-Wei Lee; Tristan Tay; Kanae Sasaki; Vincent M Rotello

doi:10.1021/acsnano.6b07600

Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein for Efficient Gene Editing

ACS Nano. 2017 Mar 28;11(3):2452-2458. doi: 10.1021/acsnano.6b07600. Epub 2017 Jan 31.

Authors

Rubul Mout¹, Moumita Ray¹, Gulen Yesilbag Tonga¹, Yi-Wei Lee¹, Tristan Tay¹, Kanae Sasaki¹, Vincent M Rotello¹

Affiliation

¹ Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.

Abstract

Genome editing through the delivery of CRISPR/Cas9-ribonucleoprotein (Cas9-RNP) reduces unwanted gene targeting and avoids integrational mutagenesis that can occur through gene delivery strategies. Direct and efficient delivery of Cas9-RNP into the cytosol followed by translocation to the nucleus remains a challenge. Here, we report a remarkably highly efficient (∼90%) direct cytoplasmic/nuclear delivery of Cas9 protein complexed with a guide RNA (sgRNA) through the coengineering of Cas9 protein and carrier nanoparticles. This construct provides effective (∼30%) gene editing efficiency and opens up opportunities in studying genome dynamics.

Keywords: CRISPR delivery; CRISPR/Cas9; CRISPR/Cas9 engineering; gene editing; genome engineering; nanoparticle.

Publication types

Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

CRISPR-Cas Systems / genetics*
Cytosol / chemistry
Cytosol / metabolism*
Gene Editing*
Gene Transfer Techniques*
Protein Engineering
Ribonucleoproteins / chemistry
Ribonucleoproteins / genetics*

Substances

Ribonucleoproteins

Grants and funding

R01 GM077173/GM/NIGMS NIH HHS/United States