title: Direct Cytosolic Delivery of CRISPR/Cas9-Ribonucleoprotein for Efficient Gene Editing.
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.
authors.affiliation: Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Rubul
authors.identifier: 0000-0001-6125-5873
authors.initials: R
authors.lastname: Mout
authors.affiliation: Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Moumita
authors.initials: M
authors.lastname: Ray
authors.affiliation: Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Gulen
authors.initials: G
authors.lastname: Yesilbag Tonga
authors.affiliation: Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Yi-Wei
authors.initials: YW
authors.lastname: Lee
authors.affiliation: Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Tristan
authors.initials: T
authors.lastname: Tay
authors.affiliation: Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Kanae
authors.initials: K
authors.lastname: Sasaki
authors.affiliation: Department of Chemistry, University of Massachusetts , 710 North Pleasant Street, Amherst, Massachusetts 01003, United States.
authors.forename: Vincent M
authors.identifier: 0000-0002-5184-5439
authors.initials: VM
authors.lastname: Rotello
mesh_terms: D064113:CRISPR-Cas Systems / Q000235:genetics*; D003600:Cytosol / Q000737:chemistry / Q000378:metabolism*; D000072669:Gene Editing; D018014:Gene Transfer Techniques; D015202:Protein Engineering; D012261:Ribonucleoproteins / Q000737:chemistry / Q000235:genetics*
chemical_list: D012261:Ribonucleoproteins
keywords: CRISPR delivery; CRISPR/Cas9; CRISPR/Cas9 engineering; gene editing; genome engineering; nanoparticle