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

2017; American Chemical Society; Volume: 11; Issue: 3 Linguagem: Inglês

10.1021/acsnano.6b07600

1936-086X

Rubul Mout, Moumita Ray, Gülen Yesilbag Tonga, Yi-Wei Lee, Tristan Tay, Kanae Sasaki, Vincent M. Rotello,

Advanced biosensing and bioanalysis techniques

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.