DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins
Abstract
Editing plant genomes without introducing foreign DNA into cells may alleviate regulatory concerns related to genetically modified plants. We transfected preassembled complexes of purified Cas9 protein and guide RNA into plant protoplasts of Arabidopsis thaliana, tobacco, lettuce and rice and achieved targeted mutagenesis in regenerated plants at frequencies of up to 46%. The targeted sites contained germline-transmissible small insertions or deletions that are indistinguishable from naturally occurring genetic variation.
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Effective delivery of Cas9/sgRNA ribonucleoprotein complexes (RNPs) can be difficult due to the large size of Cas9 protein (~160 kD), the positive charge of Cas9 protein and the strong negative charge of sgRNA.
“However, due to the large size of Cas9 protein (~ 160 kD), the positive charge of Cas9 protein and the strong negative charge of sgRNA, the effective delivery of Cas9/sgRNA ribonucleoprotein complexes (RNPs) can be difficult [88–91].”
From Non-viral delivery systems for CRISPR/Cas9-based genome editing: challenges and opportunities (2018) · cites this paper as [90]
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- Authors
- Woo, J. W., Kim, J., Kwon, S. I., Corvalán, C., Cho, S. W., Kim, H., Kim, S.-G., Kim, S.-T., Choe, S., & Kim, J.-S.
- Affiliations
- Advanced Institute of Convergence Technology (KR)
- Seoul National University (KR)
- Institute for Basic Science (KR)
- Corresponding author
- Choe, Sunghwa
- Kim, Jin-Soo
- Venue
- Nature Biotechnology
- Published
- 2015-10-19
- Volume
- 33
- Issue
- 11
- Pages
- 1162-1164
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W1787401975
- Cited by
- 1,219
- References
- 23
- Citations by year
- 2026: 67 · 2025: 89 · 2024: 101 · 2023: 116 · 2022: 140 · 2021: 175 · 2020: 128 · 2019: 127 · 2018: 90 · 2017: 94 · 2016: 84 · 2015: 8
- FWCI
- 39.93
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Seoul National University Open Repository (Seoul National University) (submittedVersion)
- Scholarworks@UNIST (Ulsan National Institute of Science and Technology) (submittedVersion)
- Funders
- Institute for Basic Science
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Transgenic Plants and Applications
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Genetically Modified Organisms Research
- Keywords
- CRISPR/Cas9, DNA-free genome editing, Cas9, guide RNA, targeted mutagenesis, germline transmission, Arabidopsis thaliana, tobacco, lettuce, rice, transgenic plants, regulatory concerns
- MeSH terms
- Base Sequence, Genetic Engineering, Molecular Sequence Data, Plant Proteins, Protoplasts, Ribonucleoproteins, Oryza, Nicotiana, Arabidopsis, Genome, Plant, Plants, Genetically Modified, Gene Knockout Techniques, CRISPR-Cas Systems
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