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

Wujin Sun; Wenyan Ji; Jordan M Hall; Quanyin Hu; Chao Wang; Chase L Beisel; Zhen Gu

doi:10.1002/anie.201506030

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

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

Authors

Wujin Sun^{1

2}, Wenyan Ji^{1

2}, Jordan M Hall³, Quanyin Hu^{1

2}, Chao Wang^{1

2}, Chase L Beisel⁴, Zhen Gu^{5

6

7}

Affiliations

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). [email protected].
⁵ Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695 (USA). [email protected].
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). [email protected].
⁷ Department of Medicine, University of North Carolina School of Medicine, Chapel Hill, NC 27599 (USA). [email protected].

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.

Publication types

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

MeSH terms

Animals
CRISPR-Associated Proteins / administration & dosage*
CRISPR-Associated Proteins / chemistry
CRISPR-Associated Proteins / genetics
CRISPR-Cas Systems*
Cell Line
Clustered Regularly Interspaced Short Palindromic Repeats
DNA / administration & dosage*
DNA / chemistry
DNA / genetics
Green Fluorescent Proteins / genetics
Humans
Mice, Nude
Nanoparticles / chemistry*
Neoplasms / genetics
Neoplasms / therapy
RNA, Guide, CRISPR-Cas Systems / administration & dosage*
RNA, Guide, CRISPR-Cas Systems / chemistry
RNA, Guide, CRISPR-Cas Systems / genetics
Streptococcus pyogenes / chemistry
Streptococcus pyogenes / genetics

Substances

CRISPR-Associated Proteins
RNA, Guide, CRISPR-Cas Systems
enhanced green fluorescent protein
Green Fluorescent Proteins
DNA

Abstract

Publication types

MeSH terms

Substances

Grants and funding