Engineered CRISPR-Cas9 nuclease with expanded targeting space
Open accessAbstract
Cas9 enzyme (SpCas9) requires an NGG protospacer adjacent motif (PAM) for target recognition, thereby restricting the targetable genomic loci. Here, we report a rationally engineered SpCas9 variant (SpCas9-NG) that can recognize relaxed NG PAMs. The crystal structure revealed that the loss of the base-specific interaction with the third nucleobase is compensated by newly introduced non-base-specific interactions, thereby enabling the NG PAM recognition. We showed that SpCas9-NG induces indels at endogenous target sites bearing NG PAMs in human cells. Furthermore, we found that the fusion of SpCas9-NG and the activation-induced cytidine deaminase (AID) mediates the C-to-T conversion at target sites with NG PAMs in human cells.
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SpCas9-NG and SpG can target NGN PAMs.
“These can be targeted in combination with evolved Cas9 variants such as SpCas9-NG20 or SpG.19”
From Protocol to create isogenic disease models from adult stem cell-derived organoids using next-generation CRISPR tools (2024) · cites this paper as [20]
The paper reports that SpCas9-NG edited sites with NGA, NGT, NGG, and NGC PAMs, although activity was lower at NGC sites (section labelled Abstract; page not provided). Our copy does not mention SpG, so it supports only the SpCas9-NG part of the claim. (LLM, conf=0.88)
Experimental — accuracy not yet measuredA rationally engineered SpCas9 variant (SpCas9-NG) generated based on the reduced interaction between the third nucleobase of PAMs and the gRNA-Cas9 complex could edit the target site with NG PAMs.
“A rationally engineered SpCas9 variant (SpCas9-NG) generated based on the reduced interaction between the third nucleobase of PAMs and the gRNA-Cas9 complex could edit the target site with NG PAMs.”
From Machine learning-based prediction models to guide the selection of Cas9 variants for efficient gene editing (2024) · cites this paper as [28]
Cas9 genomic localization is delimited by the PAM, which is NGG for SpCas9 while evolved variants of SpCas9 tolerate a broader range of PAM sequences.
“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 [19]
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<i>Coot</i> : model-building tools for molecular graphics
Emsley, P., & Cowtan, K. (2004) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444904019158 · Cited by 30,571 · see citing works
<i>PHENIX</i> : a comprehensive Python-based system for macromolecular structure solution
Adams, P. D., Afonine, P. V., Bunkóczi, G., Chen, V. B., Davis, I. W., Echols, N., Headd, J. J., Hung, L.-W., Kapral, G. J., Grosse-Kunstleve, R. W., et al. (2010) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444909052925 · Cited by 23,716 · see citing works
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 · see citing works
Kabsch, W. (2010) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444909047337 · Cited by 16,463 · see citing works
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 · see citing works
Self-citationProgrammable editing of a target base in genomic DNA without double-stranded DNA cleavage
Komor, A. C., Kim, Y. B., Packer, M. S., Zuris, J. A., & Liu, D. R. (2016) · Nature · doi:10.1038/nature17946 · Cited by 5,437 · see citing works
Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System
Zetsche, B., Gootenberg, J. S., Abudayyeh, O. O., Slaymaker, I. M., Makarova, K. S., Essletzbichler, P., Volz, S. E., Joung, J., John van der, O. O. S. T., Regev, A., et al. (2015) · Cell · doi:10.1016/j.cell.2015.09.038 · Cited by 4,750 · see citing works
Self-citationHow good are my data and what is the resolution?
Evans, P. R., & Murshudov, G. N. (2013) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444913000061 · Cited by 4,747 · see citing works
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 · see citing works
Self-citationMolecular replacement with MOLREP
Vagin, A. A., & Teplyakov, A. (2009) · Acta Crystallographica Section D Biological Crystallography · doi:10.1107/s0907444909042589 · Cited by 3,580 · see citing works
In vivo genome editing using Staphylococcus aureus Cas9
Ran, F. A., Cong, L., Yan, W. X., Scott, D. A., Gootenberg, J. S., Kriz, A. J., Zetsche, B., Shalem, O., Wu, X., Makarova, K. S., et al. (2015) · Nature · doi:10.1038/nature14299 · Cited by 2,687 · see citing works
Self-citationRationally engineered Cas9 nucleases with improved specificity
Slaymaker, I. M., Gao, L., Zetsche, B., Scott, D. A., Yan, W. X., & Zhang, F. (2016) · Science · doi:10.1126/science.aad5227 · Cited by 2,414 · see citing works
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Tsai, S. Q., Zheng, Z., Nguyen, N. T., Liebers, M., Topkar, V. V., Thapar, V., Wyvekens, N., Khayter, C., Iafrate, A. J., Le, L. P., et al. (2015) · Nature Biotechnology · doi:10.1038/nbt.3117 · Cited by 2,268 · see citing works
DNA interrogation by the CRISPR RNA-guided endonuclease Cas9
Sternberg, S. H., Redding, S., Jínek, M., Greene, E. C., & Doudna, J. A. (2014) · Nature · doi:10.1038/nature13011 · Cited by 1,775 · see citing works
Engineered CRISPR-Cas9 nucleases with altered PAM specificities
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., et al. (2015) · Nature · doi:10.1038/nature14592 · Cited by 1,624 · see citing works
Evolved Cas9 variants with broad PAM compatibility and high DNA specificity
Hu, J. H., Miller, S. M., Geurts, M. H., Tang, W., Chen, L., Sun, N., Zeina, C. M., Gao, X., Rees, H. A., Lin, Z., et al. (2018) · Nature · doi:10.1038/nature26155 · Cited by 1,552 · see citing works
Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems
Nishida, K., Arazoe, T., Yachie, N., Banno, S., Kakimoto, M., Tabata, M., Mochizuki, M., Miyabe, A., Araki, M., Hara, K. Y., et al. (2016) · Science · doi:10.1126/science.aaf8729 · Cited by 1,349 · see citing works
Self-citationStructural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease
Anders, C., Niewoehner, O., Duerst, A., & Jínek, M. (2014) · Nature · doi:10.1038/nature13579 · Cited by 1,319 · see citing works
Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage
Jiang, F., Taylor, D. W., Chen, J. S., Kornfeld, J. E., Zhou, K., Thompson, A. J., Nogales, E., & Doudna, J. A. (2016) · Science · doi:10.1126/science.aad8282 · Cited by 616 · see citing works
Engineered Cpf1 variants with altered PAM specificities
Gao, L., Cox, D. B. T., Yan, W. X., Manteiga, J. C., Schneider, M. W., Yamano, T., Nishimasu, H., Nureki, O., Crosetto, N., & Zhang, F. (2017) · Nature Biotechnology · doi:10.1038/nbt.3900 · Cited by 460 · see citing works
Self-citationCrystal Structure of Staphylococcus aureus Cas9
Nishimasu, H., Cong, L., Yan, W. X., Ran, F. A., Zetsche, B., Li, Y., Kurabayashi, A., Ishitani, R., Zhang, F., & Nureki, O. (2015) · Cell · doi:10.1016/j.cell.2015.08.007 · Cited by 430 · see citing works
Self-citationStructural Basis for the Canonical and Non-canonical PAM Recognition by CRISPR-Cpf1
Yamano, T., Zetsche, B., Ishitani, R., Zhang, F., Nishimasu, H., & Nureki, O. (2017) · Molecular Cell · doi:10.1016/j.molcel.2017.06.035 · Cited by 302 · see citing works
Self-citationStructural Plasticity of PAM Recognition by Engineered Variants of the RNA-Guided Endonuclease Cas9
Anders, C., Bargsten, K., & Jinek, M. (2016) · Molecular Cell · doi:10.1016/j.molcel.2016.02.020 · Cited by 201 · see citing works
Structural basis for the altered PAM specificities of engineered CRISPR-cas9
Hirano, S., Nishimasu, H., Ishitani, R., & Nureki, O. (2016) · Molecular Cell · doi:10.1016/j.molcel.2016.02.018 · Cited by 152 · see citing works
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Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors
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Base editing: precision chemistry on the genome and transcriptome of living cells
Rees, H. A., & Liu, D. R. (2018) · Nature Reviews Genetics · doi:10.1038/s41576-018-0059-1 · Cited by 1,675 · see citing works
CRISPR/Cas Genome Editing and Precision Plant Breeding in Agriculture
Chen, K., Wang, Y., Zhang, R., Zhang, H., & Gao, C. (2019) · Annual Review of Plant Biology · doi:10.1146/annurev-arplant-050718-100049 · Cited by 1,643 · see citing works
Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants
Walton, R. T., Christie, K. A., Whittaker, M. N., & Kleinstiver, B. P. (2020) · Science · doi:10.1126/science.aba8853 · Cited by 1,233 · see citing works
Richter, M. F., Zhao, K. T., Eton, E., Lapinaite, A., Newby, G. A., Thuronyi, B. W., Wilson, C., Koblan, L. W., Zeng, J., Bauer, D. E., et al. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0453-z · Cited by 1,020 · see citing works
Applications of CRISPR–Cas in agriculture and plant biotechnology
Zhu, H., Li, C., & Gao, C. (2020) · Nature Reviews Molecular Cell Biology · doi:10.1038/s41580-020-00288-9 · Cited by 870 · see citing works
CRISPR/Cas9 therapeutics: progress and prospects
Li, T., Yang, Y., Qi, H., Cui, W., Zhang, L., Fu, X., He, X., Liu, M., Li, P., & Yu, T. (2023) · Signal Transduction and Targeted Therapy · doi:10.1038/s41392-023-01309-7 · Cited by 770 · see citing works
Prime editing for precise and highly versatile genome manipulation
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Past, present, and future of CRISPR genome editing technologies
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CRISPR Gene Therapy: Applications, Limitations, and Implications for the Future
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CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells
Kurt, I. C., Zhou, R., Iyer, S., Garcia, S. P., Miller, B. R., Langner, L. M., Grünewald, J., & Joung, J. K. (2021) · Nature Biotechnology · doi:10.1038/s41587-020-0609-x · Cited by 573 · see citing works
CRISPR/Cas system: recent advances and future prospects for genome editing
Manghwar, H., Lindsey, K., Zhang, X., & Jin, S. (2019) · Trends in Plant Science · doi:10.1016/j.tplants.2019.09.006 · Cited by 529 · see citing works
Directed evolution of adenine base editors with increased activity and therapeutic application
Gaudelli, N. M., Lam, D. K., Rees, H. A., Solá-Esteves, N. M., Barrera, L. A., Born, D. A., Edwards, A., Gehrke, J. M., Lee, S., Liquori, A., et al. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0491-6 · Cited by 518 · see citing works
CRISPR-Cas9 DNA Base-Editing and Prime-Editing
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Chai, A. C., Cui, M., Chemello, F., Li, H. Z., Chen, K., Tan, W. H., Atmanli, A., McAnally, J., Zhang, Y., Xu, L., et al. (2023) · Nature Medicine · doi:10.1038/s41591-022-02176-5 · Cited by 187 · see citing works
Genome editing in plants by engineered CRISPR–Cas9 recognizing NG PAM
Endo, M., Mikami, M., Endo, A., Kaya, H., Itoh, T., Nishimasu, H., Nureki, O., & Toki, S. (2018) · Nature Plants · doi:10.1038/s41477-018-0321-8 · Cited by 185 · see citing works
Self-citationDevelopment of CRISPR-Cas systems for genome editing and beyond
Zhang, F. (2019) · Quarterly Reviews of Biophysics · doi:10.1017/s0033583519000052 · Cited by 185 · see citing works
Self-citationJang, H., Jo, D. H., Cho, C. S., Shin, J. H., Seo, J. H., Yu, G., Gopalappa, R., Kim, D., Cho, S., Kim, J. H., et al. (2021) · Nature Biomedical Engineering · doi:10.1038/s41551-021-00788-9 · Cited by 184 · see citing works
Bastet, A., Zafirov, D., Giovinazzo, N., Guyon‐Debast, A., Nogué, F., Robaglia, C., & Gallois, J. (2019) · Plant Biotechnology Journal · doi:10.1111/pbi.13096 · Cited by 182 · see citing works
Zhang, B. (2020) · Journal of Cellular Physiology · doi:10.1002/jcp.30064 · Cited by 182 · see citing works
Yeh, W.-H., Shubina-Oleinik, O., Levy, J. M., Pan, B., Newby, G. A., Wornow, M., Burt, R., Chen, J. C., Holt, J. R., & Liu, D. R. (2020) · Science Translational Medicine · doi:10.1126/scitranslmed.aay9101 · Cited by 179 · see citing works
CRISPR–Cas: a tool for cancer research and therapeutics
Yin, H., Xue, W., & Anderson, D. G. (2019) · Nature Reviews Clinical Oncology · doi:10.1038/s41571-019-0166-8 · Cited by 178 · see citing works
Precision genome editing using cytosine and adenine base editors in mammalian cells
Huang, T. P., Newby, G. A., & Liu, D. R. (2021) · Nature Protocols · doi:10.1038/s41596-020-00450-9 · Cited by 175 · see citing works
A catalogue of biochemically diverse CRISPR-Cas9 orthologs
Gasiūnas, G., Young, J. K., Karvelis, T., Kazlauskas, D., Urbaitis, T., Jasnauskaitė, M., Grusyte, M. M., Paulraj, S., Wang, P.-H., Hou, Z., et al. (2020) · Nature Communications · doi:10.1038/s41467-020-19344-1 · Cited by 175 · see citing works
Perspectives on the Application of Genome-Editing Technologies in Crop Breeding
Hua, K., Zhang, J., Botella, J. R., Ma, C., Kong, F., Liu, B., & Zhu, J. (2019) · Molecular Plant · doi:10.1016/j.molp.2019.06.009 · Cited by 170 · see citing works
CRISPR/Cas: Advances, Limitations, and Applications for Precision Cancer Research
Yang, Y., Xu, J., Ge, S., & Lai, L. (2021) · Frontiers in Medicine · doi:10.3389/fmed.2021.649896 · Cited by 160 · see citing works
CRISPR-Cas9/Cas12a biotechnology and application in bacteria
Yao, R., Liu, D., Jia, X., Zheng, Y., Liu, W. Y., & Xiao, Y. (2018) · Synthetic and Systems Biotechnology · doi:10.1016/j.synbio.2018.09.004 · Cited by 159 · see citing works
Chen, L., Zhu, B., Ru, G., Meng, H., Yan, Y., Hong, M., Zhang, D., Luan, C., Zhang, S., Wu, H., et al. (2023) · Nature Biotechnology · doi:10.1038/s41587-022-01532-7 · Cited by 156 · see citing works
Current Status and Challenges of DNA Base Editing Tools
Jeong, Y. K., Song, B., & Bae, S. (2020) · Molecular Therapy · doi:10.1016/j.ymthe.2020.07.021 · Cited by 153 · see citing works
Evaluating CRISPR-based prime editing for cancer modeling and CFTR repair in organoids
Geurts, M. H., de Poel, E., Pleguezuelos-Manzano, C., Oka, R., Carrillo, L., Andersson-Rolf, A., Boretto, M., Brunsveld, J. E., van Boxtel, R., Beekman, J. M., et al. (2021) · Life Science Alliance · doi:10.26508/lsa.202000940 · Cited by 151 · see citing works
Ren, B., Liu, L., Li, S., Kuang, Y., Wang, J., Zhang, D., Zhou, X., Lin, H., & Zhou, H. (2019) · Molecular Plant · doi:10.1016/j.molp.2019.03.010 · Cited by 146 · see citing works
Kim, H. K., Lee, S., Kim, Y., Park, J., Min, S., Choi, J. W., Huang, T. P., Yoon, S., Liu, D. R., & Kim, H. H. (2020) · Nature Biomedical Engineering · doi:10.1038/s41551-019-0505-1 · Cited by 145 · see citing works
Exploration of Plant-Microbe Interactions for Sustainable Agriculture in CRISPR Era
Shelake, R. M., Pramanik, D., & Kim, J. (2019) · Microorganisms · doi:10.3390/microorganisms7080269 · Cited by 141 · see citing works
RNA‐based medicine: from molecular mechanisms to therapy
Sparmann, A., & Vogel, J. (2023) · The EMBO Journal · doi:10.15252/embj.2023114760 · Cited by 141 · see citing works
Genome Engineering in Rice Using Cas9 Variants that Recognize NG PAM Sequences
Hua, K., Tao, X., Han, P., Wang, R., & Zhu, J. (2019) · Molecular Plant · doi:10.1016/j.molp.2019.03.009 · Cited by 140 · see citing works
The Caenorhabditis elegans Transgenic Toolbox
Nance, J., & Frøkjær‐Jensen, C. (2019) · Genetics · doi:10.1534/genetics.119.301506 · Cited by 140 · see citing works
Zhang, X., Chen, L., Zhu, B., Wang, L., Chen, C., Hong, M., Huang, Y., Li, H., Han, H., Cai, B., et al. (2020) · Nature Cell Biology · doi:10.1038/s41556-020-0518-8 · Cited by 140 · see citing works
High-efficiency and multiplex adenine base editing in plants using new TadA variants
Yan, D., Ren, B., Liu, L., Yan, F., Li, S., Wang, G., Sun, W., Zhou, X., & Zhou, H. (2021) · Molecular Plant · doi:10.1016/j.molp.2021.02.007 · Cited by 140 · see citing works
Base editing rescue of spinal muscular atrophy in cells and in mice
Arbab, M., Matuszek, Ż., Kray, K. M., Du, A., Newby, G. A., Blatnik, A. J., Raguram, A., Richter, M. F., Zhao, K. T., Levy, J. M., et al. (2023) · Science · doi:10.1126/science.adg6518 · Cited by 140 · see citing works
CRISPR–Cas applications in agriculture and plant research
Tuncel, A., Pan, C., Clem, J. S., Liu, D., & Qi, Y. (2025) · Nature Reviews Molecular Cell Biology · doi:10.1038/s41580-025-00834-3 · Cited by 139 · see citing works
Peptide-mediated delivery of CRISPR enzymes for the efficient editing of primary human lymphocytes
Foss, D. V., Muldoon, J. J., Nguyen, D. N., Carr, D., Sahu, S. U., Hunsinger, J. M., Wyman, S. K., Krishnappa, N., Mendonsa, R., Schanzer, E. V., et al. (2023) · Nature Biomedical Engineering · doi:10.1038/s41551-023-01032-2 · Cited by 138 · see citing works
Kumar, D., Yadav, A., Ahmad, R., Dwivedi, U. N., & Yadav, K. (2022) · Frontiers in Genetics · doi:10.3389/fgene.2022.932859 · Cited by 132 · see citing works
Improved plant cytosine base editors with high editing activity, purity, and specificity
Ren, Q., Sretenovic, S., Liu, G., Zhong, Z., Wang, J., Huang, L., Tang, X., Guo, Y., Liu, L., Wu, Y., et al. (2021) · Plant Biotechnology Journal · doi:10.1111/pbi.13635 · Cited by 131 · see citing works
Chen, L., Hong, M., Luan, C., Gao, H., Ru, G., Guo, X., Zhang, D., Zhang, S., Li, C., Wu, J., et al. (2023) · Nature Biotechnology · doi:10.1038/s41587-023-01821-9 · Cited by 131 · see citing works
Hino, T., Omura, S. N., Nakagawa, R., Togashi, T., Takeda, S. N., Hiramoto, T., Tasaka, S., Hirano, H., Tokuyama, T., Uosaki, H., et al. (2023) · Cell · doi:10.1016/j.cell.2023.08.031 · Cited by 131 · see citing works
Self-citationVersatile detection with CRISPR/Cas system from applications to challenges
Wu, H., Chen, X., Zhang, M., Wang, X., Chen, Y., Qian, C., Wu, J. F., & Xu, J. (2020) · TrAC Trends in Analytical Chemistry · doi:10.1016/j.trac.2020.116150 · Cited by 130 · see citing works
An engineered ScCas9 with broad PAM range and high specificity and activity
Chatterjee, P., Jakimo, N., Lee, J., Amrani, N., Rodríguez, T., Koseki, S., Tysinger, E., Qing, R., Hao, S., Sontheimer, E. J., et al. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0517-0 · Cited by 128 · see citing works
Prediction of efficiencies for diverse prime editing systems in multiple cell types
Yu, G., Kim, H. K., Park, J., Kwak, H., Cheong, Y., Kim, D., Kim, J.-Y., Kim, J., & Kim, H. H. (2023) · Cell · doi:10.1016/j.cell.2023.03.034 · Cited by 128 · see citing works
Genome editing in large animals: current status and future prospects
Zhao, J., Lai, L., Ji, W., & Zhou, Q. (2019) · National Science Review · doi:10.1093/nsr/nwz013 · Cited by 126 · see citing works
Dead Cas Systems: Types, Principles, and Applications
Brezgin, S., Kostyusheva, A., Kostyushev, D. S., & Chulanov, V. (2019) · International Journal of Molecular Sciences · doi:10.3390/ijms20236041 · Cited by 126 · see citing works
Massively parallel kinetic profiling of natural and engineered CRISPR nucleases
Jones, S. K., Hawkins, J. A., Johnson, N. V., Jung, C., Hu, K., Rybarski, J. R., Chen, J. S., Doudna, J. A., Press, W. H., & Finkelstein, I. J. (2020) · Nature Biotechnology · doi:10.1038/s41587-020-0646-5 · Cited by 126 · see citing works
CRISPR/dCas9-Based Systems: Mechanisms and Applications in Plant Sciences
Karlson, C. K. S., Noor, S. N. M., Nolte, N., & Tan, B. C. (2021) · Plants · doi:10.3390/plants10102055 · Cited by 126 · see citing works
The Solanum tuberosum GBSSI gene: a target for assessing gene and base editing in tetraploid potato
Veillet, F., Chauvin, L., Kermarrec, M.-P., Sevestre, F., Merrer, M., Terret, Z., Szydlowski, N., Devaux, P., Gallois, J.-L., & Chauvin, J.-E. (2019) · Plant Cell Reports · doi:10.1007/s00299-019-02426-w · Cited by 124 · see citing works
Expanding the CRISPR Toolbox in Zebrafish for Studying Development and Disease
Liu, K., Petree, C., Requena, T., Varshney, P., & Varshney, G. K. (2019) · Frontiers in Cell and Developmental Biology · doi:10.3389/fcell.2019.00013 · Cited by 124 · see citing works
Base editing in plants: Current status and challenges
Bharat, S. S., Li, S., Li, J., Yan, L., & Xia, L. (2019) · The Crop Journal · doi:10.1016/j.cj.2019.10.002 · Cited by 123 · see citing works
Massively parallel base editing to map variant effects in human hematopoiesis
Martin-Rufino, J. D., Castano, N., Pang, M., Grody, E. I., Joubran, S. R., Caulier, A., Wahlster, L., Li, T., Qiu, X., Riera-Escandell, A. M., et al. (2023) · Cell · doi:10.1016/j.cell.2023.03.035 · Cited by 123 · see citing works
A Cas9 with PAM recognition for adenine dinucleotides
Chatterjee, P., Lee, J., Nip, L., Koseki, S., Tysinger, E., Sontheimer, E. J., Jacobson, J. M., & Jakimo, N. (2020) · Nature Communications · doi:10.1038/s41467-020-16117-8 · Cited by 121 · see citing works
- Authors
- Nishimasu, H., Shi, X., Ishiguro, S., Gao, L., Hirano, S., Okazaki, S., Noda, T., Abudayyeh, O. O., Gootenberg, J. S., Mori, H., Oura, S., Holmes, B., Tanaka, M., Seki, M., Hirano, H., Aburatani, H., Ishitani, R., Ikawa, M., Yachie, N., Zhang, F., & Nureki, O.
- Affiliations
- The University of Tokyo (JP)
- Broad Institute (US)
- Keio University (JP)
- Research Institute for Microbial Diseases, The University of Osaka (JP)
- Research Center for Advanced Science and Technology, The University of Tokyo (JP)
- Institute of Medical Science, The University of Tokyo (JP)
- Corresponding author
- Nishimasu, Hiroshi
- Nureki, Osamu
- ORCID
- Venue
- Science
- Published
- 2018-09-21
- Volume
- 361
- Issue
- 6408
- Pages
- 1259-1262
- ISSN
- 0036-8075 (ISSN-L)
- 1095-9203
- PMC
- PMC6368452
- OpenAlex
- W2889513238
- Cited by
- 1,100
- References
- 25
- Citations by year
- 2026: 81 · 2025: 113 · 2024: 143 · 2023: 134 · 2022: 167 · 2021: 169 · 2020: 156 · 2019: 125 · 2018: 11
- FWCI
- 41.07
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Science (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- DSpace@MIT (Massachusetts Institute of Technology) (submittedVersion, cc-by-nc-sa)
- PMC
- Funders
- Japan Agency for Medical Research and Development
- 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, crystal structure, indels, C-to-T conversion, activation-induced cytidine deaminase, targeted mutagenesis
- MeSH terms
- Gene Editing, CRISPR-Associated Protein 9, Bacterial Proteins, Endonucleases, Humans, Protein Engineering, Crystallography, X-Ray, HEK293 Cells, CRISPR-Cas Systems
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