The DOI matches its Crossref record; 36 of the 36 references it cites were found in a registry or an index.
Engineered CRISPR-Cas9 nucleases with altered PAM specificities
Open accessAbstract
Although CRISPR-Cas9 nucleases are widely used for genome editing, the range of sequences that Cas9 can recognize is constrained by the need for a specific protospacer adjacent motif (PAM). As a result, it can often be difficult to target double-stranded breaks (DSBs) with the precision that is necessary for various genome-editing applications. The ability to engineer Cas9 derivatives with purposefully altered PAM specificities would address this limitation. Here we show that the commonly used Streptococcus pyogenes Cas9 (SpCas9) can be modified to recognize alternative PAM sequences using structural information, bacterial selection-based directed evolution, and combinatorial design. These altered PAM specificity variants enable robust editing of endogenous gene sites in zebrafish and human cells not currently targetable by wild-type SpCas9, and their genome-wide specificities are comparable to wild-type SpCas9 as judged by GUIDE-seq analysis. In addition, we identify and characterize another SpCas9 variant that exhibits improved specificity in human cells, possessing better discrimination against off-target sites with non-canonical NAG and NGA PAMs and/or mismatched spacers. We also find that two smaller-size Cas9 orthologues, Streptococcus thermophilus Cas9 (St1Cas9) and Staphylococcus aureus Cas9 (SaCas9), function efficiently in the bacterial selection systems and in human cells, suggesting that our engineering strategies could be extended to Cas9s from other species. Our findings provide broadly useful SpCas9 variants and, more importantly, establish the feasibility of engineering a wide range of Cas9s with altered and improved PAM specificities.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Precision editing of alleles in a human cell has drastically improved the understanding of genotypic and phenotypic effects in gene therapy due to the lower off-target rates [119,120,121].”
From Utility of iPSC-Derived Cells for Disease Modeling, Drug Development, and Cell Therapy (2022) · cites this paper as [120]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Besides, there are other variants that could identify different PAM sequences, such as VQR, VRER, VRQR, and QQR1, allowing gene editing on a wider range of genetic loci.”
From Machine learning-based prediction models to guide the selection of Cas9 variants for efficient gene editing (2024) · cites this paper as [25]
Experimental — accuracy not yet measuredThe cited source says what the claim attributes to it.
In the citing paper
“The basic bacterial selection scheme performed was adapted from a previously described positive selection system for identifying Cas9 mutants in E. coli31.”
From A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells (2018) · cites this paper as [31]
The cited source says what the claim attributes to it.
In the citing paper
“To be applicable to a wider range of genomic sites, some non-NGG PAM Cas9 were generated.”
From Machine learning-based prediction models to guide the selection of Cas9 variants for efficient gene editing (2024) · cites this paper as [25]
The cited source says what the claim attributes to it.
In the citing paper
“This could result in the development of wider-ranged VQR, EQR, and VRER SpCas9 variants, enabling the targeting of NGAN, NGNG, and NGCG PAMs, respectively (Kleinstiver et al., 2015; Hirano et al., 2016).”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Kleinstiver et al., 2015
The cited source says what the claim attributes to it.
In the citing paper
“These modifications have led to the creation of SpCas9 variants such as VQR, EQR, and VRER, which can recognize broader PAM sequences like NGAN, NGNG, and NGCG, respectively (Kleinstiver et al., 2015; Hirano et al., 2016).”
From Emerging applications of gene editing technologies for the development of climate-resilient crops (2025) · cites this paper as Kleinstiver et al., 2015
The cited source says what the claim attributes to it.
In the citing paper
“Cas9 genomic localization is delimited by the protospacer adjacent motif (PAM), which is NGG for SpCas9 while evolved variants of SpCas9 tolerate a broader range of PAM sequences17–21.”
From One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids (2023) · cites this paper as [18]
The check wasn’t confident either way, so it gives no verdict — never disputed.
In the citing paper
“This screen identified four key beneficial point mutations in SpCas9, giving rise to evoCas9, a variant that exhibits fidelity exceeding SpCas9-HF1 and eSpCas9 (Kleinstiver et al., 2015; Casini et al., 2018).”
From Off-target effects in CRISPR/Cas9 gene editing (2023) · cites this paper as Kleinstiver et al., 2015
Experimental — accuracy not yet measuredThe check wasn’t confident either way, so it gives no verdict — never disputed.
In the citing paper
“SaKKHcas9 is an evolved variant of SaCas9 and recognizes the more relaxed NNNRRT PAM18.”
From One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids (2023) · cites this paper as [18]
Experimental — accuracy not yet measured
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
Fast and accurate short read alignment with Burrows-Wheeler transform
Li, H., & Durbin, R. (2009) · Bioinformatics · doi:10.1093/bioinformatics/btp324 · Cited by 58,607
Initial sequencing and analysis of the human genome
International Human Genome Sequencing Consortium, Whitehead Institute for Biomedical Research, C. F. G. R., Lander, E. S., Linton, L. M., Birren, B., Nusbaum, C., Zody, M. C., Baldwin, J., Devon, K., Dewar, K., et al. (2001) · Nature · doi:10.1038/35057062 · Cited by 24,216
A Programmable Dual-RNA–Guided DNA Endonuclease in Adaptive Bacterial Immunity
Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J. A., & Charpentier, E. (2012) · Science · doi:10.1126/science.1225829 · Cited by 17,116
Multiplex Genome Engineering Using CRISPR/Cas Systems
Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P. D., Wu, X., Jiang, W., Marraffini, L. A., et al. (2013) · Science · doi:10.1126/science.1231143 · Cited by 15,339
RNA-Guided Human Genome Engineering via Cas9
Mali, P., Yang, L., Esvelt, K. M., Aach, J., Guell, M., DiCarlo, J. E., Norville, J. E., & Church, G. M. (2013) · Science · doi:10.1126/science.1232033 · Cited by 9,210
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
DNA targeting specificity of RNA-guided Cas9 nucleases
Hsu, P. D., Scott, D. A., Weinstein, J. A., Ran, F. A., Konermann, S., Agarwala, V., Li, Y., Fine, E. J., Wu, X., Shalem, O., et al. (2013) · Nature Biotechnology · doi:10.1038/nbt.2647 · Cited by 4,741
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
High-Frequency off-Target Mutagenesis Induced by CRISPR-Cas Nucleases in Human Cells
Fu, Y., Foden, J. A., Khayter, C., Maeder, M. L., Reyon, D., Joung, J. K., & Sander, J. D. (2013) · Nature Biotechnology · doi:10.1038/nbt.2623 · Cited by 3,292
Self-citationCRISPR-Cas systems for editing, regulating and targeting genomes
Sander, J. D., & Joung, J. K. (2014) · Nature Biotechnology · doi:10.1038/nbt.2842 · Cited by 3,153
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Items 401–500 of 1,001
CRISPR/Cas9-based genome editing in the era of CAR T cell immunotherapy
Salas-McKee, J., Kong, W., Gladney, W. L., Jadlowsky, J. K., Plesa, G., Suhoski, M. M., & Fraietta, J. A. (2019) · Human Vaccines & Immunotherapeutics · doi:10.1080/21645515.2019.1571893 · Cited by 68
Considerations in adapting CRISPR/Cas9 in nongenetic model plant systems
Shan, S., Soltis, P. S., Soltis, D. E., & Yang, B. (2020) · Applications in Plant Sciences · doi:10.1002/aps3.11314 · Cited by 71
Utility of iPSC-Derived Cells for Disease Modeling, Drug Development, and Cell Therapy
Nicholson, M. W., Ting, C.-Y., Chan, D. Z. H., Cheng, Y.-C., Lee, Y.-C., Hsu, C.-C., Huang, C.-Y., & Hsieh, P. C. H. (2022) · Cells · doi:10.3390/cells11111853 · Cited by 65
One-step generation of tumor models by base editor multiplexing in adult stem cell-derived organoids
Geurts, M. H., Gandhi, S., Boretto, M. G., Akkerman, N., Derks, L. L. M., van Son, G., Celotti, M., Harshuk-Shabso, S., Peci, F., Begthel, H., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-40701-3 · Cited by 66
Efficient CRISPR-mediated base editing in Agrobacterium spp.
Rodrigues, S. D., Karimi, M., Impens, L., Lerberge, E. V., Coussens, G., Aesaert, S., Rombaut, D., Holtappels, D., Ibrahim, H. M. M., MONTAGU, M. V., et al. (2020) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.2013338118 · Cited by 66
CRISPR/Cas Technology in Pig-to-Human Xenotransplantation Research
Ryczek, N., Hryhorowicz, M., Zeyland, J., Lipiński, D., & Słomski, R. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms22063196 · Cited by 66
High-throughput functional evaluation of human cancer-associated mutations using base editors
Kim, Y., Lee, S., Cho, S., Park, J., Chae, D., Park, T., Minna, J. D., & Kim, S. (2022) · Nature Biotechnology · doi:10.1038/s41587-022-01276-4 · Cited by 66
Common variation in atrial fibrillation: navigating the path from genetic association to mechanism
Tucker, N. R., Clauß, S., & Ellinor, P. T. (2016) · Cardiovascular Research · doi:10.1093/cvr/cvv283 · Cited by 66
Lei, C., Li, S., Liu, J.-K., Zheng, X., Zhao, G., & Wang, J. (2017) · Nucleic Acids Research · doi:10.1093/nar/gkx018 · Cited by 66
Gene editing and its applications in biomedicine
Li, G., Li, X., Zhuang, S., Wang, L., Zhu, Y., Chen, Y., Sun, W. C., Wu, Z., Zhou, Z. L., Chen, J., et al. (2022) · Science China Life Sciences · doi:10.1007/s11427-021-2057-0 · Cited by 66
Recent Advances in Improving Gene-Editing Specificity through CRISPR–Cas9 Nuclease Engineering
Huang, X., Yang, D., Zhang, J., Xu, J., & Chen, Y. E. (2022) · Cells · doi:10.3390/cells11142186 · Cited by 68
Engineered Nanomaterials to Potentiate CRISPR/Cas9 Gene Editing for Cancer Therapy
Yi, K., Kong, H., Lao, Y., Li, D., Mintz, R. L., Fang, T., Chen, G., Tao, Y., Li, M., & Ding, J. (2023) · Advanced Materials · doi:10.1002/adma.202300665 · Cited by 66
Evolution of CRISPR/Cas Systems for Precise Genome Editing
Hryhorowicz, M., Lipiński, D., & Zeyland, J. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms241814233 · Cited by 69
Developing a highly efficient and wildly adaptive CRISPR ‐SaCas9 toolset for plant genome editing
Qin, R., Li, J., Li, H., Zhang, Y., Liu, X., Miao, Y., Zhang, X., & Wei, P. (2018) · Plant Biotechnology Journal · doi:10.1111/pbi.13047 · Cited by 65
Structural insights into a high fidelity variant of SpCas9
Guo, M., Ren, K., Zhu, Y., Tang, Z., Wang, Y., Zhang, B., & Huang, Z. (2019) · Cell Research · doi:10.1038/s41422-018-0131-6 · Cited by 65
Applying CRISPR–Cas9 tools to identify and characterize transcriptional enhancers
Lopes, R., Korkmaz, G., & Agami, R. (2016) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm.2016.79 · Cited by 65
Engineer chimeric Cas9 to expand PAM recognition based on evolutionary information
Ma, D., Xu, Z., Zhang, Z., Chen, X., Zeng, X., Zhang, Y., Deng, T., Ren, M., Sun, Z., Jiang, R., et al. (2019) · Nature Communications · doi:10.1038/s41467-019-08395-8 · Cited by 66
Genome-wide CRISPR-Cas9 screening in mammalian cells
Yu, J., & Yusa, K. (2019) · Methods · doi:10.1016/j.ymeth.2019.04.015 · Cited by 66
Stukenberg, D., Hoff, J., Faber, A., & Becker, A. (2022) · Communications Biology · doi:10.1038/s42003-022-03150-0 · Cited by 67
DNA methylation and de-methylation using hybrid site-targeting proteins
Lei, Y., Huang, Y., & Goodell, M. A. (2018) · Genome biology · doi:10.1186/s13059-018-1566-2 · Cited by 67
Wang, Y., Liu, Y., Li, J., Yang, Y., Ni, X., Cheng, H., Huang, T., Guo, Y., Ma, H., Zheng, P., et al. (2019) · Biotechnology and Bioengineering · doi:10.1002/bit.27121 · Cited by 64
Maggio, I., Zittersteijn, H. A., Wang, Q., Liu, J., Janssen, J. M., Ojeda, I. T., van der Maarel, S. M., Lankester, A. C., Hoeben, R. C., & Gonçalves, M. A. F. V. (2020) · Gene Therapy · doi:10.1038/s41434-019-0119-y · Cited by 66
The compact Casπ (Cas12l) ‘bracelet’ provides a unique structural platform for DNA manipulation
(2023) · doi:10.1038/s41422-022-00771-2 · Cited by 64
Self-citationBenchmarking of SpCas9 variants enables deeper base editor screens of BRCA1 and BCL2
Sangree, A. K., Griffith, A. L., Szegletes, Z. M., Roy, P., DeWeirdt, P. C., Hegde, M., McGee, A. V., Hanna, R. E., & Doench, J. G. (2022) · Nature Communications · doi:10.1038/s41467-022-28884-7 · Cited by 64
Kim, H. S., Kweon, J., & Kim, Y. (2024) · Experimental & Molecular Medicine · doi:10.1038/s12276-024-01212-3 · Cited by 68
Advances in detecting and reducing off-target effects generated by CRISPR-mediated genome editing
Li, J., Hong, S., Chen, W., Zuo, E., & Yang, H. (2019) · Journal of genetics and genomics/Journal of Genetics and Genomics · doi:10.1016/j.jgg.2019.11.002 · Cited by 66
O’Hagan, S., & Kell, D. B. (2015) · PeerJ · doi:10.7717/peerj.1405 · Cited by 64
CRISPR-Cas9 Gene Editing for Fruit and Vegetable Crops: Strategies and Prospects
Wan, L., Wang, Z., Tang, M., Hong, D., Sun, Y., Ren, J. M., Zhang, N., & Zeng, H. (2021) · Horticulturae · doi:10.3390/horticulturae7070193 · Cited by 64
CRISPR-Cas gene editing technology and its application prospect in medicinal plants
Guo, M., Chen, H., Dong, S., Zhang, Z., & Luo, H. (2022) · Chinese Medicine · doi:10.1186/s13020-022-00584-w · Cited by 66
Unraveling the mechanisms of PAMless DNA interrogation by SpRY-Cas9
Hibshman, G. N., Bravo, J. P. K., Hooper, M. M., Dangerfield, T. L., Zhang, H., Finkelstein, I. J., Johnson, K. A., & Taylor, D. W. (2024) · Nature Communications · doi:10.1038/s41467-024-47830-3 · Cited by 70
Genome Editing with CRISPR-Cas9: Can It Get Any Better?
Haeussler, M., & Concordet, J. (2016) · Journal of genetics and genomics/Journal of Genetics and Genomics · doi:10.1016/j.jgg.2016.04.008 · Cited by 65
Barakate, A., & Stephens, J. (2016) · Frontiers in Plant Science · doi:10.3389/fpls.2016.00765 · Cited by 64
Efficient In Vivo Liver-Directed Gene Editing Using CRISPR/Cas9
Singh, K., Evens, H., Nair, N., Rincón, M. Y., Sarcar, S., Samara-Kuko, E., Chuah, M., & VandenDriessche, T. (2018) · Molecular Therapy · doi:10.1016/j.ymthe.2018.02.023 · Cited by 63
Zhao, C., Zheng, X., Qu, W., Li, G., Li, X., Miao, Y., Han, X., Liu, X., Li, Z., Ma, Y., et al. (2017) · International Journal of Biological Sciences · doi:10.7150/ijbs.21312 · Cited by 62
Self-citationMotta, B. M., Pramstaller, P. P., Hicks, A. A., & Rossini, A. (2017) · Stem Cells International · doi:10.1155/2017/8960236 · Cited by 64
Enhancing gene editing specificity by attenuating DNA cleavage kinetics
Miller, J. C., Patil, D. P., Xia, D. F., Paine, C. B., Fauser, F., Richards, H. W., Shivak, D. A., Bendaña, Y. R., Hinkley, S. J., Scarlott, N. A., et al. (2019) · Nature Biotechnology · doi:10.1038/s41587-019-0186-z · Cited by 63
Efficient generation of Knock-in/Knock-out marmoset embryo via CRISPR/Cas9 gene editing
Kumita, W., Sato, K., Suzuki, Y., Kurotaki, Y., Harada, T., Zhou, Y., Kishi, N., Sato, K., Aiba, A., Sakakibara, Y., et al. (2019) · Scientific Reports · doi:10.1038/s41598-019-49110-3 · Cited by 64
Emerging Species and Genome Editing Tools: Future Prospects in Cyanobacterial Synthetic Biology
Gale, G. A. R., Schiavon Osorio, A. A., Mills, L. A., Wang, B., Lea-Smith, D. J., & McCormick, A. J. (2019) · Microorganisms · doi:10.3390/microorganisms7100409 · Cited by 61
Kelley, C. P., Haerle, M. C., & Wang, E. T. (2022) · Cell Reports · doi:10.1016/j.celrep.2022.111226 · Cited by 61
Effective knockdown of Drosophila long non-coding RNAs by CRISPR interference
Ghosh, S., Tibbit, C., & Liu, J. (2016) · Nucleic Acids Research · doi:10.1093/nar/gkw063 · Cited by 62
CRISPR/Cas genome editing to optimize pharmacologically active plant natural products
Dey, A. (2020) · Pharmacological Research · doi:10.1016/j.phrs.2020.105359 · Cited by 62
Deep mutational scanning of S. pyogenes Cas9 reveals important functional domains
Spencer, J. M., & Zhang, X. (2017) · Scientific Reports · doi:10.1038/s41598-017-17081-y · Cited by 65
CRISPR/Cascade 9-Mediated Genome Editing-Challenges and Opportunities
Roy, B., Zhao, J. C., Yang, C., Luo, W., Xiong, T., Li, Y., Fang, X., Gao, G., Singh, C. O., Madsen, L., et al. (2018) · Frontiers in Genetics · doi:10.3389/fgene.2018.00240 · Cited by 61
SpG and SpRY variants expand the CRISPR toolbox for genome editing in zebrafish
Liang, F., Zhang, Y., Li, L., Yang, Y., Fei, J., Liu, Y., & Qin, W. (2022) · Nature Communications · doi:10.1038/s41467-022-31034-8 · Cited by 61
Essential RNA-Based Technologies and Their Applications in Plant Functional Genomics
Teotia, S., Singh, D., Tang, X., & Tang, G. (2016) · Trends in biotechnology · doi:10.1016/j.tibtech.2015.12.001 · Cited by 60
Mammalian Synthetic Biology: Engineering Biological Systems
Black, J. B., Pérez‐Piñera, P., & Gersbach, C. A. (2017) · Annual Review of Biomedical Engineering · doi:10.1146/annurev-bioeng-071516-044649 · Cited by 60
PAM-flexible genome editing with an engineered chimeric Cas9
Zhao, L., Koseki, S. R. T., Silverstein, R. A., Amrani, N., Peng, C., Kramme, C., Savic, N., Pacesa, M., Rodríguez, T. C., Stan, T., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-41829-y · Cited by 59
Self-citationCurrent trends of clinical trials involving CRISPR/Cas systems
Zhang, S., Wang, Y., Mao, D., Wang, Y., Zhang, H., Pan, Y., Wang, Y., Teng, S., & Huang, P. (2023) · Frontiers in Medicine · doi:10.3389/fmed.2023.1292452 · Cited by 62
CRISPR-Cas9 for medical genetic screens: applications and future perspectives
Xue, H.-Y., Ji, L.-J., Gao, A.-M., Liu, P., He, J.-D., & Lu, X. (2015) · Journal of Medical Genetics · doi:10.1136/jmedgenet-2015-103409 · Cited by 62
Idoko-Akoh, A., Taylor, L., Sang, H., & McGrew, M. J. (2018) · Scientific Reports · doi:10.1038/s41598-018-33244-x · Cited by 62
Swinnen, G., Meyer, M. D., Pollier, J., Molina‐Hidalgo, F. J., Ceulemans, E., Venegas‐Molina, J., Milde, L. D., Fernández‐Calvo, P., Ron, M., Pauwels, L., et al. (2022) · New Phytologist · doi:10.1111/nph.18379 · Cited by 59
Nanobiomaterial vectors for improving gene editing and gene therapy
Hu, S. W., Ding, T., Tang, H., Guo, H., Cui, W., & Shu, Y. (2023) · Materials Today · doi:10.1016/j.mattod.2023.04.011 · Cited by 58
Integrating machine learning and genome editing for crop improvement
Chen, L., Liu, G., & Zhang, T. (2024) · aBIOTECH · doi:10.1007/s42994-023-00133-5 · Cited by 60
Epigenome editing for targeted DNA (de)methylation: a new perspective in modulating gene expression
Seem, K., Kaur, S., Kumar, S., & Mohapatra, T. (2024) · Critical Reviews in Biochemistry and Molecular Biology · doi:10.1080/10409238.2024.2320659 · Cited by 59
Hu, J. H., Davis, K. M., & Liu, D. R. (2016) · Cell chemical biology · doi:10.1016/j.chembiol.2015.12.009 · Cited by 61
Rational Design of Mini-Cas9 for Transcriptional Activation
Ma, D., Peng, S., Huang, W., Cai, Z., & Xie, Z. (2018) · ACS Synthetic Biology · doi:10.1021/acssynbio.7b00404 · Cited by 59
CRISPR-based gene expression control for synthetic gene circuits
Santos‐Moreno, J., & Schaerli, Y. (2020) · Biochemical Society Transactions · doi:10.1042/bst20200020 · Cited by 59
Schmidt, M. J., Gupta, A. K., Bednarski, C., Gehrig-Giannini, S., Richter, F. M., Pitzler, C., Gamalinda, M., Galonska, C., Takeuchi, R., Wang, K., et al. (2021) · Nature Communications · doi:10.1038/s41467-021-24454-5 · Cited by 58
Konstantinidis, E., Molisak, A., Perrin, F., Streubel-Gallasch, L., Fayad, S., Kim, D. Y., Petri, K., Aryee, M. J., Aguilar, X., György, B., et al. (2022) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2022.03.022 · Cited by 60
Self-citationMasarwy, R., Stotsky‐Oterin, L., Elisha, A., Hazan‐Halevy, I., & Peer, D. (2024) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2024.115359 · Cited by 59
Chaikind, B., Bessen, J. L., Thompson, D. B., Hu, J. H., & Liu, D. R. (2016) · Nucleic Acids Research · doi:10.1093/nar/gkw707 · Cited by 57
Integration and exchange of split dCas9 domains for transcriptional controls in mammalian cells
Ma, D., Peng, S., & Xie, Z. (2016) · Nature Communications · doi:10.1038/ncomms13056 · Cited by 57
Robust genome editing of CRISPR-Cas9 at NAG PAMs in rice
Meng, X., Hu, X., Liu, Q., Song, X., Gao, C., Li, J., & Wang, K. (2017) · Science China Life Sciences · doi:10.1007/s11427-017-9247-9 · Cited by 57
Klompe, S. E., & Sternberg, S. H. (2018) · The CRISPR Journal · doi:10.1089/crispr.2018.0012 · Cited by 57
CRISPR/Cas mediated epigenome editing for cancer therapy
Ansari, I., Chaturvedi, A., Chitkara, D., & Singh, S. P. (2021) · Seminars in Cancer Biology · doi:10.1016/j.semcancer.2020.12.018 · Cited by 58
A CRISPR Path to Engineering New Genetic Mouse Models for Cardiovascular Research
Miano, J. M., Zhu, Q. M., & Lowenstein, C. J. (2016) · Arteriosclerosis Thrombosis and Vascular Biology · doi:10.1161/atvbaha.116.304790 · Cited by 58
The Conspicuity of CRISPR-Cpf1 System as a Significant Breakthrough in Genome Editing
Bayat, H., Modarressi, M. H., & Rahimpour, A. (2017) · Current Microbiology · doi:10.1007/s00284-017-1406-8 · Cited by 56
CRISPR-Based Tools in Immunity
Simeonov, D. R., & Marson, A. (2019) · Annual Review of Immunology · doi:10.1146/annurev-immunol-042718-041522 · Cited by 57
Directed Evolution of CRISPR/Cas Systems for Precise Gene Editing
Liu, R., Liang, L., Freed, E. F., & Gill, R. T. (2020) · Trends in biotechnology · doi:10.1016/j.tibtech.2020.07.005 · Cited by 58
Martella, A., Firth, M. A., Taylor, B., Göppert, A., Cuomo, E. M., Roth, R. G., Dickson, A. J., & Fisher, D. I. (2019) · ACS Synthetic Biology · doi:10.1021/acssynbio.8b00527 · Cited by 56
CRISPR/Cas Genome Editing—Can It Become a Game Changer in Future Fisheries Sector?
Roy, S., Kumar, V., Behera, B. K., Parhi, J., Mohapatra, S., Chakraborty, T., & Das, B. K. (2022) · Frontiers in Marine Science · doi:10.3389/fmars.2022.924475 · Cited by 55
DNA base editing in nuclear and organellar genomes
Tan, J., Forner, J., Karcher, D., & Bock, R. (2022) · Trends in Genetics · doi:10.1016/j.tig.2022.06.015 · Cited by 57
CRISPR/Cas9 Gene Editing: An Unexplored Frontier for Forest Pathology
Dort, E. N., Tanguay, P., & Hamelin, R. C. (2020) · Frontiers in Plant Science · doi:10.3389/fpls.2020.01126 · Cited by 57
CRISPR-Cas systems: ushering in the new genome editing era
Rojo, F. P., Nyman, R. K. M., Johnson, A. A. T., Pazos-Navarro, M., Ryan, M. H., Erskine, W., & Kaur, P. (2018) · Bioengineered · doi:10.1080/21655979.2018.1470720 · Cited by 58
Efficient plant genome engineering using a probiotic sourced CRISPR-Cas9 system
Zhong, Z., Liu, G., Tang, Z., Xiang, S., Yang, L., Huang, L., He, Y., Fan, T., Liu, S., Zheng, X., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-41802-9 · Cited by 58
Wolfs, J. M., Hamilton, T. A., Lant, J. T., Laforet, M., Zhang, J., Salemi, L. M., Gloor, G. B., Schild‐Poulter, C., & Edgell, D. R. (2016) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1616343114 · Cited by 56
Heler, R., Wright, A. V., Vucelja, M., Bikard, D., Doudna, J. A., & Marraffini, L. A. (2016) · Molecular Cell · doi:10.1016/j.molcel.2016.11.031 · Cited by 57
Genome editing for inborn errors of metabolism: advancing towards the clinic
Schneller, J. L., Lee, C. M., Bao, G., & Venditti, C. P. (2017) · BMC Medicine · doi:10.1186/s12916-017-0798-4 · Cited by 57
Advancing Metabolic Engineering of Saccharomyces cerevisiae Using the CRISPR/Cas System
Lian, J., HamediRad, M., & Zhao, H. (2018) · Biotechnology Journal · doi:10.1002/biot.201700601 · Cited by 54
Targeted Nucleotide Editing Technologies for Microbial Metabolic Engineering
Arazoe, T., Kondo, A., & Nishida, K. (2018) · Biotechnology Journal · doi:10.1002/biot.201700596 · Cited by 54
Fei, J., Lou, W. P.-K., Knapp, D., Murawala, P., Gerber, T., Taniguchi, Y., Nowoshilow, S., Khattak, S., & Tanaka, E. M. (2018) · Nature Protocols · doi:10.1038/s41596-018-0071-0 · Cited by 57
Mendoza, B. J., & Trinh, C. T. (2017) · Bioinformatics · doi:10.1093/bioinformatics/btx564 · Cited by 55
Guyon, A., Rousseau, J., Bégin, F.-G., Bertin, T., Lamothe, G., & Tremblay, J. P. (2021) · Molecular Therapy — Nucleic Acids · doi:10.1016/j.omtn.2021.02.032 · Cited by 55
CRISPR mediated genome engineering to develop climate smart rice: Challenges and opportunities
Biswal, A. K., Mangrauthia, S. K., Reddy, M. R., & Yugandhar, P. (2019) · Seminars in Cell and Developmental Biology · doi:10.1016/j.semcdb.2019.04.005 · Cited by 56
CRISPR/Cas9 for cancer treatment: technology, clinical applications and challenges
Cheng, X., Fan, S., Wen, C., & Du, X. (2020) · Briefings in Functional Genomics · doi:10.1093/bfgp/elaa001 · Cited by 55
Detection and Modulation of DNA Translocations During Multi-Gene Genome Editing in T Cells
Bothmer, A., Gareau, K. W., Abdulkerim, H. S., Buquicchio, F. A., Cohen, L. A., Viswanathan, R., Zuris, J. A., Marco, E., Fernández, C. A., Myer, V. E., et al. (2020) · The CRISPR Journal · doi:10.1089/crispr.2019.0074 · Cited by 56
History of genome editing: From meganucleases to CRISPR
Tröder, S. E., & Zevnik, B. (2021) · Laboratory Animals · doi:10.1177/0023677221994613 · Cited by 55
Prokaryotic Argonaute Proteins as a Tool for Biotechnology
Kropocheva, E., Lisitskaya, L., Agapov, A., Musabirov, A. A., Kulbachinskiy, A. V., & Esyunina, D. (2022) · Molecular Biology · doi:10.1134/s0026893322060103 · Cited by 54
Genetic Renal Diseases: The Emerging Role of Zebrafish Models
Elmonem, M. A., Berlingerio, S. P., van den Heuvel, L. P., de Witte, P. A. M., Lowe, M., & Levtchenko, E. N. (2018) · Cells · doi:10.3390/cells7090130 · Cited by 59
Repurposing the Streptococcus mutans CRISPR-Cas9 System to Understand Essential Gene Function
Shields, R. C., Walker, A. R., Maricic, N., Chakraborty, B., Underhill, S. A. M., & Burne, R. A. (2020) · PLoS Pathogens · doi:10.1371/journal.ppat.1008344 · Cited by 52
Sretenovic, S., Yin, D., Levav, A., Selengut, J., Mount, S. M., & Qi, Y. (2020) · Plant Communications · doi:10.1016/j.xplc.2020.100101 · Cited by 53
CRISPR_Cas systems for fungal research
Ouedraogo, J., & Tsang, A. (2020) · Fungal Biology Reviews · doi:10.1016/j.fbr.2020.10.002 · Cited by 54
Aðalsteinsson, B. Þ., Kristjansdottir, T., Merré, W., Helleux, A., Dusaucy, J., Tourigny, M., Friðjónsson, Ó. H., & Hreggviðsson, G. Ó. (2021) · Scientific Reports · doi:10.1038/s41598-021-89029-2 · Cited by 56
Recent Progress and Challenges in CRISPR-Cas9 Engineered Algae and Cyanobacteria
Patel, V. K., Das, A., Kumari, R., & Kajla, S. (2023) · Algal Research · doi:10.1016/j.algal.2023.103068 · Cited by 50
Jeong, Y. K., Yu, J., & Bae, S. (2019) · Scientific Reports · doi:10.1038/s41598-019-41356-1 · Cited by 51
Rittiner, J. E., Moncalvo, M., Chiba‐Falek, O., & Kantor, B. (2020) · Frontiers in Molecular Neuroscience · doi:10.3389/fnmol.2020.00148 · Cited by 53
Reading Frame Repair of TTN Truncation Variants Restores Titin Quantity and Functions
Romano, R., Ghahremani, S., Zimmerman, T., Legere, N., Thakar, K., Ladha, F. A., Pettinato, A. M., & Hinson, J. T. (2021) · Circulation · doi:10.1161/circulationaha.120.049997 · Cited by 52
Precise DNA cleavage using CRISPR-SpRYgests
Christie, K. A., Guo, J. A., Silverstein, R. A., Doll, R. M., Mabuchi, M. Y., Stutzman, H. E., Lin, J., Ma, L., Walton, R. T., Pinello, L., et al. (2022) · Nature Biotechnology · doi:10.1038/s41587-022-01492-y · Cited by 51
Self-citationSufyan, M., Daraz, U., Hyder, S., Zulfiqar, U., Iqbal, R., Eldin, S. M., Rafiq, F., Mahmood, N., Shahzad, K., Uzair, M., et al. (2023) · Functional & Integrative Genomics · doi:10.1007/s10142-023-01036-w · Cited by 55
Allemailem, K. S., Almatroodi, S. A., Almatroudi, A., Alrumaihi, F., Abdulmonem, W. A., Al‐Megrin, W. A. I., Aljamaan, A. N., Rahmani, A. H., & Khan, A. A. (2023) · International Journal of Molecular Sciences · doi:10.3390/ijms24087052 · Cited by 53
Metadata
- Authors
- Kleinstiver, B. P., Prew, M. S., Tsai, S. Q., Topkar, V. V., Nguyen, N. T., Zheng, Z., Gonzales, A. P. W., Li, Z., Peterson, R. T., Yeh, J.-R. J., Aryee, M. J., & Joung, J. K.
- Affiliations
- Harvard University (US)
- Massachusetts General Hospital (US)
- Broad Institute (US)
- Corresponding author
- Joung, J. Keith
- ORCID
- Venue
- Nature
- Published
- 2015-06-22
- Volume
- 523
- Issue
- 7561
- Pages
- 481-485
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC4540238
- OpenAlex
- W4245053291
- Semantic Scholar
- afe29e1767b5d0f3563ee9f2c1a586a9d5e8cb9b
- Cited by
- 1,624
- References
- 36
- Citations by year
- 2026: 58 · 2025: 105 · 2024: 117 · 2023: 134 · 2022: 166 · 2021: 165 · 2020: 164 · 2019: 154 · 2018: 147 · 2017: 186 · 2016: 167 · 2015: 58
- FWCI
- 80.95
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Digital Access to Scholarship at Harvard (DASH) (Harvard University) (submittedVersion, cc-by)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- National Institute of General Medical Sciences (DP1GM105378, R01GM107427, R01GM088040)
- National Center for Chronic Disease Prevention and Health Promotion (DP1 GM105378)
- 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
- Health Sciences › Medicine › Oncology › CAR-T cell therapy research
- Keywords
- CRISPR/Cas9, SpCas9, protospacer adjacent motif, genome editing, off-target effects, SaCas9, Cas9 nickase, zebrafish
- MeSH terms
- Animals, Cell Line, Humans, Mutation, Staphylococcus aureus, Streptococcus pyogenes, Substrate Specificity, Zebrafish, Protein Engineering, Genome, Directed Molecular Evolution, Amino Acid Substitution, Streptococcus thermophilus, Nucleotide Motifs, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems, CRISPR-Associated Proteins
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
