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Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12029-33. doi: 10.1002/anie.201506030. Epub 2015 Aug 27.

Self-assembled DNA nanoclews for the efficient delivery of CRISPR-Cas9 for genome editing.

Sun W1,2, Ji W1,2, Hall JM3, Hu Q1,2, Wang C1,2, Beisel CL4, Gu Z5,6,7.

Author information

1
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695 (USA).
2
Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 (USA).
3
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695-7905 (USA).
4
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695-7905 (USA). cbeisel@ncsu.edu.
5
Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695 (USA). zgu@email.unc.edu.
6
Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599 (USA). zgu@email.unc.edu.
7
Department of Medicine, University of North Carolina School of Medicine, Chapel Hill, NC 27599 (USA). zgu@email.unc.edu.

Abstract

CRISPR-Cas9 represents a promising platform for genome editing, yet means for its safe and efficient delivery remain to be fully realized. A novel vehicle that simultaneously delivers the Cas9 protein and single guide RNA (sgRNA) is based on DNA nanoclews, yarn-like DNA nanoparticles that are synthesized by rolling circle amplification. The biologically inspired vehicles were efficiently loaded with Cas9/sgRNA complexes and delivered the complexes to the nuclei of human cells, thus enabling targeted gene disruption while maintaining cell viability. Editing was most efficient when the DNA nanoclew sequence and the sgRNA guide sequence were partially complementary, offering a design rule for enhancing delivery. Overall, this strategy provides a versatile method that could be adapted for delivering other DNA-binding proteins or functional nucleic acids.

KEYWORDS:

CRISPR-Cas9; DNA; drug delivery; genome editing; nanoparticles

PMID:
26310292
PMCID:
PMC4677991
DOI:
10.1002/anie.201506030
[Indexed for MEDLINE]
Free PMC Article

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