CRISPR prime editing with ribonucleoprotein complexes in zebrafish and primary human cells
Open accessAbstract
Prime editors have been delivered using DNA or RNA vectors. Here we demonstrate prime editing with purified ribonucleoprotein complexes. We introduced somatic mutations in zebrafish embryos with frequencies as high as 30% and demonstrate germline transmission. We also observed unintended insertions, deletions and prime editing guide RNA (pegRNA) scaffold incorporations. In HEK293T and primary human T cells, prime editing with purified ribonucleoprotein complexes introduced desired edits with frequencies of up to 21 and 7.5%, respectively.
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Delivery of next-generation CRISPR tools by lentivirus or ribonucleoprotein complexes may be a suitable alternative strategy to DNA mutations in adult stem cells.
“Thus, delivery of next-generation CRISPR tools by lentivirus or ribonucleoprotein complexes may be a suitable alternative strategy to DNA mutations in adult stem cells.54,55,56”
From Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools (2024) · cites this paper as [54]
The Abstract reports prime editing delivered as ribonucleoprotein complexes in zebrafish embryos and primary human T cells, and suggests that RNP delivery could broaden therapeutic applications. It does not establish lentiviral delivery or suitability in adult stem cells; it describes viral-vector delivery as challenging instead. The cited paper therefore supports the RNP possibility, but not the full claim. (LLM, conf=0.86)
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- Authors
- Petri, K., Zhang, W., Ma, J., Schmidts, A., Lee, H., Horng, J. E., Kim, D. Y., Kurt, I. C., Clement, K., Hsu, J. Y., Pinello, L., Maus, M. V., Joung, J. K., & Yeh, J.-R. J.
- Affiliations
- Harvard University (US)
- Corresponding author
- Joung, J. Keith
- Yeh, Jing-Ruey Joanna
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2021-04-29
- Volume
- 40
- Issue
- 2
- Pages
- 189-193
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- PMC
- PMC8553808
- OpenAlex
- W3158544805
- Cited by
- 208
- References
- 19
- Citations by year
- 2026: 23 · 2025: 27 · 2024: 49 · 2023: 44 · 2022: 44 · 2021: 21
- FWCI
- 12.48
- Type
- journal article
- Language
- en
- Update notices
- correction (2021-05-13)
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- Deutsche Forschungsgemeinschaft (417577129)
- China Scholarship Council
- John Hansen Research Grant by the DKMS
- U.S. Department of Health & Human Services | NIH | National Human Genome Research Institute (RM1 HG009490, R35 HG010717)
- U.S. Department of Health & Human Services | NIH | National Institute of General Medical Sciences (R35 GM118158, R01 GM134069)
- United States Department of Defense | Defense Advanced Research Projects Agency (HR0011-17-2-0042)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Keywords
- ribonucleoprotein complexes, germline transmission, zebrafish embryos, human T cells, HEK293T cells, genome editing
- MeSH terms
- Gene Editing, RNA, Guide, CRISPR-Cas Systems, Animals, Humans, Ribonucleoproteins, Zebrafish, RNA, Guide, Kinetoplastida, HEK293 Cells, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems
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