The DOI matches its Crossref record; 17 of the 17 references it cites were found in a registry or an index.
Polymer-stabilized Cas9 nanoparticles and modified repair templates increase genome editing efficiency
Open accessAbstract
Versatile and precise genome modifications are needed to create a wider range of adoptive cellular therapies1–5. Here we report two improvements that increase the efficiency of CRISPR–Cas9-based genome editing in clinically relevant primary cell types. Truncated Cas9 target sequences (tCTSs) added at the ends of the homology-directed repair (HDR) template interact with Cas9 ribonucleoproteins (RNPs) to shuttle the template to the nucleus, enhancing HDR efficiency approximately two- to fourfold. Furthermore, stabilizing Cas9 RNPs into nanoparticles with polyglutamic acid further improves editing efficiency by approximately twofold, reduces toxicity, and enables lyophilized storage without loss of activity. Combining the two improvements increases gene targeting efficiency even at reduced HDR template doses, yielding approximately two to six times as many viable edited cells across multiple genomic loci in diverse cell types, such as bulk (CD3+) T cells, CD8+ T cells, CD4+ T cells, regulatory T cells (Tregs), γδ T cells, B cells, natural killer cells, and primary and induced pluripotent stem cell-derived6 hematopoietic stem progenitor cells (HSPCs). Precise genome editing is made more efficient by stabilizing Cas9 and enhancing shuttling to the nucleus.
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Citations
Cites
The new frontier of genome engineering with CRISPR-Cas9
Doudna, J. A., & Charpentier, E. (2014) · Science · doi:10.1126/science.1258096 · Cited by 7,062
Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity
Ran, F. A., Hsu, P. D., Lin, C.-Y., Gootenberg, J. S., Konermann, S., Trevino, A. E., Scott, D. A., Inoue, A., Matoba, S., Zhang, Y., et al. (2013) · Cell · doi:10.1016/j.cell.2013.08.021 · Cited by 3,299
CRISPR–Cas9 Structures and Mechanisms
Jiang, F., & Doudna, J. A. (2017) · Annual Review of Biophysics · doi:10.1146/annurev-biophys-062215-010822 · Cited by 2,029
CRISPResso2 provides accurate and rapid genome editing sequence analysis
Clement, K., Rees, H., Canver, M. C., Gehrke, J. M., Farouni, R., Hsu, J. Y., Cole, M. A., Liu, D. R., Joung, J. K., Bauer, D. E., et al. (2019) · Nature Biotechnology · doi:10.1038/s41587-019-0032-3 · Cited by 1,660
Dunbar, C. E., High, K. A., Joung, J. K., Kohn, D. B., Ozawa, K., & Sadelain, M. (2018) · Science · doi:10.1126/science.aan4672 · Cited by 1,410
Reprogramming human T cell function and specificity with non-viral genome targeting
Roth, T. L., Puig-Saus, C., Yu, R., Shifrut, E., Carnevale, J., Li, P. J., Hiatt, J., Saco, J., Krystofinski, P., Li, H., et al. (2018) · Nature · doi:10.1038/s41586-018-0326-5 · Cited by 839
Self-citationBeyond editing: repurposing CRISPR-Cas9 for precision genome regulation and interrogation
Dominguez, A. A., Lim, W. A., & Qi, L. S. (2016) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm.2015.2 · Cited by 832 · cited 1×
“CRISPR-Cas9 interacts specifically with both genomic and non-genomic dsDNA 8 , and nuclease-inactive dCas9 has been used in many applications to localize protein and RNA effectors to specific DNA sequences without cleaving the target sequence 9 .”
CIRCLE-Seq: A highly sensitive in vitro screen for genome-wide CRISPR-cas9 nuclease off-targets
Tsai, S. Q., Nguyen, N. T., Malagon-Lopez, J., Topkar, V. V., Aryee, M. J., & Joung, J. K. (2017) · Nature Methods · doi:10.1038/nmeth.4278 · Cited by 814
Vakulskas, C. A., Dever, D. P., Rettig, G. R., Turk, R., Jacobi, A. M., Collingwood, M. A., Bode, N. M., McNeill, M. S., Yan, S., Camarena, J., et al. (2018) · Nature Medicine · doi:10.1038/s41591-018-0137-0 · Cited by 782 · cited 1×
“An initial 18 cycle PCR reaction using previously validated primers 14 (see Supplementary Table 1) to an approximately 150– 250 bp region centered on the predicted gRNA cut sites.”
cGAS is activated by DNA in a length‐dependent manner
Luecke, S., Holleufer, A., Christensen, M. H., Jønsson, K. L., Boni, G. A., Sørensen, L. K., Johannsen, M., Jakobsen, M. R., Hartmann, R., & Paludan, S. R. (2017) · EMBO reports · doi:10.15252/embr.201744017 · Cited by 308
Cited by
Lipid Nanoparticle (LNP) Enables mRNA Delivery for Cancer Therapy
Zong, Y., Lin, Y., Wei, T., & Cheng, Q. (2023) · Advanced Materials · doi:10.1002/adma.202303261 · Cited by 477
Strategies in the delivery of Cas9 ribonucleoprotein for CRISPR/Cas9 genome editing
Zhang, S., Shen, J., Li, D., & Cheng, Y. (2021) · Theranostics · doi:10.7150/thno.47007 · Cited by 430
Chimeric antigen receptor natural killer (CAR-NK) cell design and engineering for cancer therapy
Gong, Y., Klein Wolterink, R. G. J., Wang, J., Bos, G. M. J., & Germeraad, W. T. V. (2021) · Journal of Hematology & Oncology · doi:10.1186/s13045-021-01083-5 · Cited by 379
Antibodies to combat viral infections: development strategies and progress
Pantaleo, G., Correia, B., Fenwick, C., Joo, V. S., & Perez, L. (2022) · Nature Reviews Drug Discovery · doi:10.1038/s41573-022-00495-3 · Cited by 279
Pooled Knockin Targeting for Genome Engineering of Cellular Immunotherapies
Roth, T. L., Li, P. J., Blaeschke, F., Nies, J. F., Apathy, R., Mowery, C., Yu, R., Nguyen, M. L. T., Lee, Y., Truong, A., et al. (2020) · Cell · doi:10.1016/j.cell.2020.03.039 · Cited by 186
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Metadata
- Authors
- Nguyen, D. N., Roth, T. L., Li, P. J., Chen, P. A., Apathy, R., Mamedov, M. R., Vo, L. T., Tobin, V. R., Goodman, D., Shifrut, E., Bluestone, J. A., Puck, J. M., Szoka, F. C., & Marson, A.
- Venue
- Nature Biotechnology
- Published
- 2019-12-09
- Volume
- 38
- Issue
- 1
- Pages
- 44-49
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W2992036306
- Cited by
- 312
- References
- 17
- FWCI
- 8.38
- Type
- journal article
- Language
- en
- Open access
- green
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- Nguyen, D. N., Roth, T. L., Li, P. J., Chen, P. A., Apathy, R., Mamedov, M. R., Vo, L. T., Tobin, V. R., Goodman, D., Shifrut, E., Bluestone, J. A., Puck, J. M., Szoka, F. C., & Marson, A. (2020). Polymer-stabilized Cas9 nanoparticles and modified repair templates increase genome editing efficiency. Nature Biotechnology. https://doi.org/10.1038/s41587-019-0325-6
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