Self‐assembled DNA nanoclews for the efficient delivery of CRISPR–Cas9 for genome editing
Open accessAbstract
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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Other methods based on nucleic acid materials, including DNA nanoclews and RNA aptamer-streptavidin complexes, were explored for delivering gene editing components.
“Other methods based on nucleic acid materials, including DNA nanoclews [234] and RNA aptamer-streptavidin complexes [235], were also explored for delivering gene editing components.”
From RNA delivery biomaterials for the treatment of genetic and rare diseases (2019) · cites this paper as [234]
Self-assembled DNA nanoclews loaded with CRISPR RNPs enabled efficient target disruption of EGFP gene in vitro and in U2OS.EGFP xenograft tumor models in vivo after intratumoral injection.
“Self-assembled DNA nanoclews were constructed for CRISPR/Cas9-based genome editing system delivery. After being loaded with CRISPR RNPs, these nanoclews enabled efficient target disruption of EGFP gene in vitro and in U2OS.EGFP xenograft tumor models in vivo after intratumoral injection [103].”
From Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018) · cites this paper as [103]
First obstacle: effective encapsulation of editing tools in delivery vectors is hindered by large size and different charge properties of Cas9 protein, mRNA and donor DNA.
“The first obstacle is effective encapsulation of the editing tools in delivery vectors due to the large size and different charge properties of Cas9 protein, mRNA and donor DNA.”
From Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018) · cites this paper as [103]
Native Cas9 protein is positively charged, while conventional proteins are negatively charged.
“Meanwhile, native Cas9 protein is positively charged, while the conventional proteins are negatively charged [88, 97, 103, 139].”
From Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018) · cites this paper as [103]
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References 39 · Cited by 629
- Authors
- Sun, W., Ji, W., Hall, J. M., Hu, Q., Wang, C., Beisel, C. L., & Gu, Z.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- North Carolina State University (US)
- Corresponding author
- Beisel, Chase L.
- Gu, Zhen
- Venue
- Angewandte Chemie International Edition
- Publisher
- Wiley
- Published
- 2015-08-27
- Volume
- 54
- Issue
- 41
- Pages
- 12029-12033
- ISSN
- 1433-7851 (ISSN-L)
- 1521-3773
- PMC
- PMC4677991
- OpenAlex
- W2140022328
- Cited by
- 629
- References
- 39
- Citations by year
- 2026: 23 · 2025: 40 · 2024: 41 · 2023: 63 · 2022: 74 · 2021: 75 · 2020: 91 · 2019: 63 · 2018: 80 · 2017: 42 · 2016: 34 · 2015: 2 · 2012: 1
- FWCI
- 22.88
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Angewandte Chemie International Edition (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- NC TraCS
- NIH (1L1TR001111)
- NSF (MCB-1452902)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced biosensing and bioanalysis techniques
- Keywords
- CRISPR/Cas9 delivery, genome editing, rolling circle amplification, single guide RNA, Cas9, DNA nanoparticles, nuclear delivery, gene disruption
- MeSH terms
- RNA, Guide, CRISPR-Cas Systems, Animals, Cell Line, DNA, Humans, Mice, Nude, Neoplasms, Streptococcus pyogenes, RNA, Guide, Kinetoplastida, Green Fluorescent Proteins, Nanoparticles, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems, CRISPR-Associated Proteins
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