Self‐Assembled DNA Nanoclews for the Efficient Delivery of CRISPR–Cas9 for Genome Editing |
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| Authors: | Wujin Sun Wenyan Ji Jordan M Hall Quanyin Hu Dr Chao Wang Prof?Dr Chase L Beisel Prof?Dr Zhen Gu |
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| Affiliation: | 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 Medicine, University of North Carolina School of Medicine, Chapel Hill, NC 27599 (USA) |
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| 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. |
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| Keywords: | CRISPR– Cas9 DNA drug delivery genome editing nanoparticles |
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