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  • Review Article
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Precise plant genome editing using base editors and prime editors

Abstract

The development of CRISPR–Cas systems has sparked a genome editing revolution in plant genetics and breeding. These sequence-specific RNA-guided nucleases can induce DNA double-stranded breaks, resulting in mutations by imprecise non-homologous end joining (NHEJ) repair or precise DNA sequence replacement by homology-directed repair (HDR). However, HDR is highly inefficient in many plant species, which has greatly limited precise genome editing in plants. To fill the vital gap in precision editing, base editing and prime editing technologies have recently been developed and demonstrated in numerous plant species. These technologies, which are mainly based on Cas9 nickases, can introduce precise changes into the target genome at a single-base resolution. This Review provides a timely overview of the current status of base editors and prime editors in plants, covering both technological developments and biological applications.

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Fig. 1: Cytosine and adenine base editing in nuclear DNA.
Fig. 2: C-to-G and dual (simultaneous C-to-T and A-to-G) base editing in nuclear DNA.
Fig. 3: Base editing in organellar DNA.
Fig. 4: Base editing in RNA.
Fig. 5: Prime editing.
Fig. 6: Base editing and prime editing applications for different kinds of genetic modifications.
Fig. 7: Examples of crop improvement by base editing and prime editing.

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References

  1. Jinek, M. et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337, 816–822 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Cong, L. et al. Multiplex genome engineering using CRISPR/Cas Systems. Science 339, 819–823 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Xie, K. & Yang, Y. RNA-guided genome editing in plants using a CRISPR-Cas system. Mol. Plant 6, 1975–1983 (2013).

    Article  CAS  PubMed  Google Scholar 

  4. Molla, K. A. & Yang, Y. Predicting CRISPR/Cas9-induced mutations for precise genome editing. Trends Biotechnol. 38, 136–141 (2020).

    Article  CAS  PubMed  Google Scholar 

  5. Huang, T. K. & Puchta, H. CRISPR/Cas-mediated gene targeting in plants: finally a turn for the better for homologous recombination. Plant Cell Rep. 38, 443–453 (2019).

    Article  CAS  PubMed  Google Scholar 

  6. Zhang, Y. & Qi, Y. Diverse systems for efficient sequence insertion and replacement in precise plant genome editing. BioDesign Res. 2020, 8659064 (2020).

    Article  Google Scholar 

  7. Lu, Y. et al. Targeted, efficient sequence insertion and replacement in rice. Nat. Biotechnol. 38, 1402–1407 (2020).

    Article  CAS  PubMed  Google Scholar 

  8. Li, S. et al. Precise gene replacement in rice by RNA transcript-templated homologous recombination. Nat. Biotechnol. 37, 445–450 (2019).

    Article  CAS  PubMed  Google Scholar 

  9. Yeh, W. H., Chiang, H., Rees, H. A., Edge, A. S. B. & Liu, D. R. In vivo base editing of post-mitotic sensory cells. Nat. Commun. 9, 2184 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  10. Molla, K. A. & Yang, Y. CRISPR/Cas-mediated base editing: technical considerations and practical applications. Trends Biotechnol. 37, 1121–1142 (2019).

    Article  CAS  PubMed  Google Scholar 

  11. Komor, A. C., Kim, Y. B., Packer, M. S., Zuris, J. A. & Liu, D. R. Programmable editing of a target base in genomic DNA without double-stranded DNA cleavage. Nature 533, 420–424 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Nishida, K. et al. Targeted nucleotide editing using hybrid prokaryotic and vertebrate adaptive immune systems. Science 353, aaf8729 (2016).

    Article  PubMed  CAS  Google Scholar 

  13. Jiang, F. et al. Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage. Science 351, 867–871 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Porto, E. M., Komor, A. C., Slaymaker, I. M. & Yeo, G. W. Base editing: advances and therapeutic opportunities. Nat. Rev. Drug Discov. 19, 839–859 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Xu, W. et al. Discriminated sgRNAs-based SurroGate system greatly enhances the screening efficiency of plant base-edited cells. Mol. Plant 13, 169–180 (2019).

    Article  PubMed  CAS  Google Scholar 

  16. Chen, Y. et al. CRISPR/Cas9-mediated base-editing system efficiently generates gain-of-function mutations in Arabidopsis. Sci. China Life Sci. 60, 520–523 (2017).

    Article  CAS  PubMed  Google Scholar 

  17. Bastet, A. et al. Mimicking natural polymorphism in eIF4E by CRISPR-Cas9 base editing is associated with resistance to potyviruses. Plant Biotechnol. J. 17, 1736–1750 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Li, Z., Xiong, X., Wang, F., Liang, J. & Li, J. F. Gene disruption through base editing-induced messenger RNA missplicing in plants. N. Phytol. 222, 1139–1148 (2019).

    Article  CAS  Google Scholar 

  19. Xue, C., Zhang, H., Lin, Q., Fan, R. & Gao, C. Manipulating mRNA splicing by base editing in plants. Sci. China Life Sci. 61, 1293–1300 (2018).

    Article  CAS  PubMed  Google Scholar 

  20. Li, J., Sun, Y., Du, J., Zhao, Y. & Xia, L. Generation of targeted point mutations in rice by a modified CRISPR/Cas9 system. Mol. Plant 10, 526–529 (2017).

    Article  CAS  PubMed  Google Scholar 

  21. Lu, Y. & Zhu, J. K. Precise editing of a target base in the rice genome using a modified CRISPR/Cas9 system. Mol. Plant 10, 523–525 (2017).

    Article  CAS  PubMed  Google Scholar 

  22. Shimatani, Z. et al. Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion. Nat. Biotechnol. 35, 441–443 (2017).

    Article  CAS  PubMed  Google Scholar 

  23. Zong, Y. et al. Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. Nat. Biotechnol. 36, 950–953 (2018).

    Article  CAS  Google Scholar 

  24. Hua, K., Tao, X. & Zhu, J. K. Expanding the base editing scope in rice by using Cas9 variants. Plant Biotechnol. J. 17, 499–504 (2019).

    Article  PubMed  Google Scholar 

  25. Ren, Q. et al. PAM-less plant genome editing using a CRISPR–SpRY toolbox. Nat. Plants 7, 25–33 (2021).

    Article  CAS  PubMed  Google Scholar 

  26. Zong, Y. et al. Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat. Biotechnol. 35, 438–440 (2017).

    Article  CAS  PubMed  Google Scholar 

  27. Zhang, R. et al. Generation of herbicide tolerance traits and a new selectable marker in wheat using base editing. Nat. Plants 5, 480–485 (2019).

    Article  CAS  PubMed  Google Scholar 

  28. Veillet, F. et al. Transgene-free genome editing in tomato and potato plants using Agrobacterium-mediated delivery of a CRISPR/Cas9 cytidine base editor. Int. J. Mol. Sci. 20, 402 (2019).

    Article  PubMed Central  CAS  Google Scholar 

  29. Veillet, F. et al. Expanding the CRISPR toolbox in P. Patens using SpCas9-NG variant and application for gene and base editing in solanaceae crops. Int. J. Mol. Sci. 21, 1024 (2020).

    Article  CAS  PubMed Central  Google Scholar 

  30. Hunziker, J. et al. Multiple gene substitution by Target-AID base-editing technology in tomato. Sci. Rep. 10, 20471 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Veillet, F. et al. The Solanum tuberosum GBSSI gene: a target for assessing gene and base editing in tetraploid potato. Plant Cell Rep. 38, 1065–1080 (2019).

    Article  CAS  PubMed  Google Scholar 

  32. Tian, S. et al. Engineering herbicide-resistant watermelon variety through CRISPR/Cas9-mediated base-editing. Plant Cell Rep. 37, 1353–1356 (2018).

    Article  CAS  PubMed  Google Scholar 

  33. Qin, L. et al. High-efficient and precise base editing of C•G to T•A in the allotetraploid cotton (Gossypium hirsutum) genome using a modified CRISPR/Cas9 system. Plant Biotechnol. J. 18, 45–56 (2020).

    Article  CAS  PubMed  Google Scholar 

  34. Cai, Y. et al. Target base editing in soybean using a modified CRISPR/Cas9 system. Plant Biotechnol. J. 18, 1996–1998 (2020).

    Article  PubMed Central  Google Scholar 

  35. Malabarba, J. et al. New strategies to overcome present CRISPR/Cas9 limitations in apple and pear: efficient dechimerization and base editing. Int. J. Mol. Sci. 22, 319 (2021).

    Article  CAS  Google Scholar 

  36. Xing, S. et al. Fine-tuning sugar content in strawberry. Genome Biol. 21, 230 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Guyon-Debast, A. et al. A blue-print for gene function analysis through base editing in the model plant Physcomitrium (Physcomitrella) patens. New Phytol. 230, 1258–1272 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Li, G., Sretenovic, S., Eisenstein, E., Coleman, G. & Qi, Y. Highly efficient C-to-T and A-to-G base editing in a Populus hybrid. Plant Biotechnol. J. 19, 1086–1088 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Wu, J. et al. Engineering herbicide-resistant oilseed rape by CRISPR/Cas9-mediated cytosine base-editing. Plant Biotechnol. J. 18, 1857–1859 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Cheng, H. et al. Base editing with high efficiency in allotetraploid oilseed rape by A3A-PBE system. Plant Biotechnol. J. 19, 87–97 (2021).

    Article  CAS  PubMed  Google Scholar 

  41. Li, X. et al. Base editing with a Cpf1-cytidine deaminase fusion. Nat. Biotechnol. 36, 324–327 (2018).

    Article  CAS  PubMed  Google Scholar 

  42. Rees, H. A. & Liu, D. R. Base editing: precision chemistry on the genome and transcriptome of living cells. Nat. Rev. Genet. 19, 770–788 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Anzalone, A. V., Koblan, L. W. & Liu, D. R. Genome editing with CRISPR–Cas nucleases, base editors, transposases and prime editors. Nat. Biotechnol. 38, 824–844 (2020).

    Article  CAS  PubMed  Google Scholar 

  44. Gehrke, J. M. et al. An APOBEC3A-Cas9 base editor with minimized bystander and off-target activities. Nat. Biotechnol. 36, 977–982 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Kim, Y. B. et al. Increasing the genome-targeting scope and precision of base editing with engineered Cas9-cytidine deaminase fusions. Nat. Biotechnol. 35, 371–376 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Liu, Z. et al. Precise base editing with CC context-specificity using engineered human APOBEC3G-nCas9 fusions. BMC Biol. 18, 111 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Li, J. et al. Genome editing mediated by SpCas9 variants with broad non-canonical protospacer-adjacent motif compatibility in plants. Mol. Plant 14, 352–360 (2021).

    Article  CAS  PubMed  Google Scholar 

  48. Zeng, D. et al. PhieCBEs: plant high-efficiency cytidine base editors with expanded target range. Mol. Plant 13, 1666–1669 (2020).

    Article  CAS  PubMed  Google Scholar 

  49. Jin, S. et al. Rationally designed APOBEC3B cytosine base editors with improved specificity. Mol. Cell 79, 728–740 (2020).

    Article  CAS  PubMed  Google Scholar 

  50. Ren, B. et al. Improved base editor for efficiently inducing genetic variations in rice with CRISPR/Cas9-guided hyperactive hAID mutant. Mol. Plant 11, 623–626 (2018).

    Article  CAS  PubMed  Google Scholar 

  51. Yu, Y. et al. Cytosine base editors with minimized unguided DNA and RNA off-target events and high on-target activity. Nat. Commun. 11, 2052 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  52. Xu, W. et al. Multiplex nucleotide editing by high-fidelity Cas9 variants with improved efficiency in rice. BMC Plant Biol. 19, 511 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Tang, X. et al. Single transcript unit CRISPR 2.0 systems for robust Cas9 and Cas12a mediated plant genome editing. Plant Biotechnol. J. 17, 1431–1445 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Endo, M. et al. Genome editing in plants by engineered CRISPR–Cas9 recognizing NG PAM. Nat. Plants 5, 14–17 (2019).

    Article  CAS  PubMed  Google Scholar 

  55. Zhang, C. et al. Expanding the base editing scope to GA and relaxed NG PAM sites by improved xCas9 system. Plant Biotechnol. J. 18, 884–886 (2020).

    Article  PubMed  Google Scholar 

  56. Sretenovic, S. et al. Expanding plant genome-editing scope by an engineered iSpyMacCas9 system that targets A-rich PAM sequences. Plant Commun. 2, 100101 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  57. Thuronyi, B. W. et al. Continuous evolution of base editors with expanded target compatibility and improved activity. Nat. Biotechnol. 37, 1070–1079 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  58. Koblan, L. W. et al. Improving cytidine and adenine base editors by expression optimization and ancestral reconstruction. Nat. Biotechnol. 36, 843–848 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Wang, M. et al. Optimizing base editors for improved efficiency and expanded editing scope in rice. Plant Biotechnol. J. 17, 1697–1699 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  60. Molla, K. A., Karmakar, S. & Islam, M. T. In CRISPR-Cas Methods. Springer Protocols Handbooks (eds Islam, M.T. et al.) (Humana, 2020); https://doi.org/10.1007/978-1-0716-0616-2_1

  61. Qin, R. et al. Developing a highly efficient and wildly adaptive CRISPR-SaCas9 toolset for plant genome editing. Plant Biotechnol. J. 18, 706–708 (2020).

    Article  CAS  Google Scholar 

  62. Wang, M. et al. Targeted base editing in rice with CRISPR/ScCas9 system. Plant Biotechnol. J. 18, 1645–1647 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  63. Wu, Y. et al. Increasing cytosine base editing scope and efficiency with engineered Cas9-PMCDA1 fusions and the modified sgRNA in rice. Front. Genet. 10, 379 (2019).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  64. Veillet, F., Kermarrec, M. P., Chauvin, L., Chauvin, J. E. & Nogué, F. CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato. PLoS ONE 15, e0235942 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  65. Chatterjee, P. et al. A Cas9 with PAM recognition for adenine dinucleotides. Nat. Commun. 11, 2474 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  66. Ren, Q. et al. Improved plant cytosine base editors with high editing activity, purity, and specificity. Plant Biotechnol. J. https://doi.org/10.1111/pbi.13635 (2021).

  67. Miller, S. M. et al. Continuous evolution of SpCas9 variants compatible with non-G PAMs. Nat. Biotechnol. 38, 471–481 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  68. Zhang, C. et al. Expanding base editing scope to near-PAMless with engineered CRISPR-Cas9 variants in plant. Mol. Plant 14, 191–194 (2020).

    Article  PubMed  CAS  Google Scholar 

  69. Nishimasu, H. et al. Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science 361, 1259–1262 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  70. Hu, J. H. et al. Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556, 57–63 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  71. Hua, K., Tao, X., Han, P., Wang, R. & Zhu, J. K. Genome engineering in rice using Cas9 variants that recognize NG PAM sequences. Mol. Plant 12, 1003–1014 (2019).

    Article  CAS  PubMed  Google Scholar 

  72. Ren, B. et al. Cas9-NG greatly expands the targeting scope of the genome-editing toolkit by recognizing NG and other atypical PAMs in rice. Mol. Plant 12, 1015–1026 (2019).

    Article  CAS  PubMed  Google Scholar 

  73. Zhong, Z. et al. Improving plant genome editing with high-fidelity xCas9 and non-canonical PAM-targeting Cas9-NG. Mol. Plant 12, 1027–1036 (2019).

    Article  CAS  PubMed  Google Scholar 

  74. Zeng, D. et al. Engineered Cas9 variant tools expand targeting scope of genome and base editing in rice. Plant Biotechnol. J. 18, 1348–1350 (2020).

    Article  PubMed  Google Scholar 

  75. Li, J. et al. Plant genome editing using xCas9 with expanded PAM compatibility. J. Genet. Genomics 46, 277–280 (2019).

    Article  PubMed  Google Scholar 

  76. Walton, R. T., Christie, K. A., Whittaker, M. N. & Kleinstiver, B. P. Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Science 368, 290–296 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  77. Xu, Z. et al. SpRY greatly expands the genome editing scope in rice with highly flexible PAM recognition. Genome Biol. 22, 6 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  78. Zetsche, B. et al. Cpf1 Is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell 163, 759–771 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  79. Zhang, Y. et al. Expanding the scope of plant genome engineering with Cas12a orthologs and highly multiplexable editing systems. Nat. Commun. 12, 1944 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  80. Kleinstiver, B. P. et al. Engineered CRISPR–Cas12a variants with increased activities and improved targeting ranges for gene, epigenetic and base editing. Nat. Biotechnol. 37, 276–282 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  81. Ming, M. et al. CRISPR–Cas12b enables efficient plant genome engineering. Nat. Plants 6, 202–208 (2020).

    Article  CAS  PubMed  Google Scholar 

  82. Zhang, X. H., Tee, L. Y., Wang, X. G., Huang, Q. S. & Yang, S. H. Off-target effects in CRISPR/Cas9-mediated genome engineering. Mol. Ther. Nucleic Acids 4, e264 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  83. Slaymaker, I. M. et al. Rationally engineered Cas9 nucleases with improved specificity. Science 351, 84–88 (2016).

    Article  CAS  PubMed  Google Scholar 

  84. Kleinstiver, B. P. et al. High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature 529, 490–495 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  85. Chen, J. S. et al. Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature 550, 407–410 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  86. Rees, H. A. et al. Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery. Nat. Commun. 8, 15790 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  87. Gao, X. et al. Treatment of autosomal dominant hearing loss by in vivo delivery of genome editing agents. Nature 553, 217–221 (2018).

    Article  CAS  PubMed  Google Scholar 

  88. Woo, J. W. et al. DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins. Nat. Biotechnol. 33, 1162–1164 (2015).

    Article  CAS  PubMed  Google Scholar 

  89. Liang, Z. et al. Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes. Nat. Commun. 8, 14261 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  90. Zuo, E. et al. Cytosine base editor generates substantial off-target single-nucleotide variants in mouse embryos. Science 364, 289–292 (2019).

    CAS  PubMed  PubMed Central  Google Scholar 

  91. Jin, S. et al. Cytosine, but not adenine, base editors induce genome-wide off-target mutations in rice. Science 364, 292–295 (2019).

    CAS  PubMed  Google Scholar 

  92. Doman, J. L., Raguram, A., Newby, G. A. & Liu, D. R. Evaluation and minimization of Cas9-independent off-target DNA editing by cytosine base editors. Nat. Biotechnol. 38, 620–628 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  93. Randall, L. B. et al. Genome- and transcriptome-wide off-target analyses of an improved cytosine base editor. Plant Physiol. kiab264 https://doi.org/10.1093/plphys/kiab264 (2021).

  94. Komor, A. C. et al. Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C:G-to-T:A base editors with higher efficiency and product purity. Sci. Adv. 3, eaao4774 (2017).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  95. Qin, R. et al. Increasing fidelity and efficiency by modifying cytidine base-editing systems in rice. Crop J. 8, 396–402 (2020).

    Article  Google Scholar 

  96. Lei, Z. et al. Detect-seq reveals out-of-protospacer editing and target-strand editing by cytosine base editors. Nat. Methods 18, 643–651 (2021).

    Article  CAS  PubMed  Google Scholar 

  97. Wang, S. et al. Precise, predictable multi-nucleotide deletions in rice and wheat using APOBEC–Cas9. Nat. Biotechnol. 38, 1460–1465 (2020).

    Article  CAS  PubMed  Google Scholar 

  98. Schiml, S., Fauser, F. & Puchta, H. The CRISPR/Cas system can be used as nuclease for in planta gene targeting and as paired nickases for directed mutagenesis in Arabidopsis resulting in heritable progeny. Plant J. 80, 1139–1150 (2014).

    Article  CAS  PubMed  Google Scholar 

  99. Wolfs, J. M. et al. Biasing genome-editing events toward precise length deletions with an RNA-guided TevCas9 dual nuclease. Proc. Natl Acad. Sci. USA 113, 14988–14993 (2016).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  100. Gaudelli, N. M. et al. Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551, 464–471 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  101. Molla, K. A., Shih, J. & Yang, Y. Single-nucleotide editing for zebra3 and wsl5 phenotypes in rice using CRISPR/Cas9-mediated adenine base editors. aBIOTECH 1, 106–118 (2020).

    Article  Google Scholar 

  102. Hua, K., Tao, X., Yuan, F., Wang, D. & Zhu, J. K. Precise A·T to G·C base editing in the rice genome. Mol. Plant 11, 627–630 (2018).

    Article  CAS  PubMed  Google Scholar 

  103. Li, C. et al. Expanded base editing in rice and wheat using a Cas9-adenosine deaminase fusion. Genome Biol. 19, 59 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  104. Yan, F. et al. Highly efficient A·T to G·C base editing by Cas9n-guided tRNA adenosine deaminase in rice. Mol. Plant 11, 631–634 (2018).

    Article  CAS  PubMed  Google Scholar 

  105. Hao, L. et al. CRISPR/Cas9-mediated adenine base editing in rice genome. Rice Science 26, 125–128 (2019).

    Article  Google Scholar 

  106. Li, J. et al. Optimizing plant adenine base editor systems by modifying the transgene selection system. Plant Biotechnol. J. 18, 1495–1497 (2020).

    Article  PubMed  Google Scholar 

  107. Kang, B.-C. et al. Precision genome engineering through adenine base editing in plants. Nat. Plants 4, 427–431 (2018).

    Article  CAS  PubMed  Google Scholar 

  108. Wang, Z. et al. ABE8e with polycistronic tRNA-gRNA expression cassette Sig-Nificantly improves adenine base editing efficiency in Nicotiana benthamiana. Int. J. Mol. Sci. 22, 5663 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  109. Yan, D. et al. High-efficiency and multiplex adenine base editing in plants using new TadA variants. Mol. Plant 14, 722–731 (2021).

    Article  CAS  PubMed  Google Scholar 

  110. Kleinstiver, B. P. et al. Engineered CRISPR-Cas9 nucleases with altered PAM specificities. Nature 523, 481–485 (2015).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  111. Negishi, K. et al. An adenine base editor with expanded targeting scope using SpCas9-NGv1 in rice. Plant Biotechnol. J. 17, 1476–1478 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  112. Ren, J. et al. Expanding the scope of genome editing with SpG and SpRY variants in rice. Sci. China Life Sci. https://doi.org/10.1007/s11427-020-1883-5 (2021).

  113. Richter, M. F. et al. Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity. Nat. Biotechnol. 38, 883–891 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  114. Grünewald, J. et al. CRISPR DNA base editors with reduced RNA off-target and self-editing activities. Nat. Biotechnol. 37, 1041–1048 (2019).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  115. Gaudelli, N. M. et al. Directed evolution of adenine base editors with increased activity and therapeutic application. Nat. Biotechnol. 38, 892–900 (2020).

    Article  CAS  PubMed  Google Scholar 

  116. Hua, K. et al. Simplified adenine base editors improve adenine base editing efficiency in rice. Plant Biotechnol. J. 18, 770–778 (2020).

    Article  CAS  PubMed  Google Scholar 

  117. Wei, C. et al. Efficient generation of homozygous substitutions in rice in one generation utilizing an rABE8e base editor. J. Integr. Plant Biol. https://doi.org/10.1111/jipb.13089 (2021).

  118. Lapinaite, A. et al. DNA capture by a CRISPR-Cas9-guided adenine base editor. Science 369, 566–571 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  119. Kim, H. S., Jeong, Y. K., Hur, J. K., Kim, J. S. & Bae, S. Adenine base editors catalyze cytosine conversions in human cells. Nat. Biotechnol. 37, 1145–1148 (2019).

    Article  CAS  PubMed  Google Scholar 

  120. Kurt, I. C. et al. CRISPR C-to-G base editors for inducing targeted DNA transversions in human cells. Nat. Biotechnol. 39, 41–46 (2021).

    Article  CAS  PubMed  Google Scholar 

  121. Zhao, D. et al. Glycosylase base editors enable C-to-A and C-to-G base changes. Nat. Biotechnol. 39, 35–40 (2021).

    Article  CAS  PubMed  Google Scholar 

  122. Chen, L. et al. Programmable C:G to G:C genome editing with CRISPR-Cas9-directed base excision repair proteins. Nat. Commun. 12, 1384 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  123. Koblan, L. W. et al. Efficient C•G-to-G•C base editors developed using CRISPRi screens, target-library analysis, and machine learning. Nat. Biotechnol. https://doi.org/10.1038/s41587-021-00938-z (2021).

  124. Molla, K. A., Qi, Y., Karmakar, S. & Baig, M. J. Base editing landscape extends to perform transversion mutation. Trends Genet. 36, 899–901 (2020).

    Article  CAS  PubMed  Google Scholar 

  125. Li, C. et al. Targeted, random mutagenesis of plant genes with dual cytosine and adenine base editors. Nat. Biotechnol. 38, 875–882 (2020).

    Article  CAS  PubMed  Google Scholar 

  126. Grünewald, J. et al. A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing. Nat. Biotechnol. 38, 861–864 (2020).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  127. Zhang, X. et al. Dual base editor catalyzes both cytosine and adenine base conversions in human cells. Nat. Biotechnol. 38, 856–860 (2020).

    Article  CAS  PubMed  Google Scholar 

  128. Sakata, R. C. et al. Base editors for simultaneous introduction of C-to-T and A-to-G mutations. Nat. Biotechnol. 38, 865–869 (2020).

    Article  CAS  PubMed  Google Scholar 

  129. Xie, J. et al. ACBE, a new base editor for simultaneous C-to-T and A-to-G substitutions in mammalian systems. BMC Biol. 18, 131 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  130. Li, C. et al. SWISS: multiplexed orthogonal genome editing in plants with a Cas9 nickase and engineered CRISPR RNA scaffolds. Genome Biol. 21, 141 (2020).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  131. Mok, B. Y. et al. A bacterial cytidine deaminase toxin enables CRISPR-free mitochondrial base editing. Nature 583, 631–637 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  132. Voytas, D. F. Plant genome engineering with sequence-specific nucleases. Annu. Rev. Plant Biol. 64, 327–350 (2013).

    Article  CAS  PubMed  Google Scholar 

  133. Aushev, M. & Herbert, M. Mitochondrial genome editing gets precise. Nature 583, 521–522 (2020).

    Article  CAS  PubMed  Google Scholar 

  134. Gualberto, J. M. & Newton, K. J. Plant mitochondrial genomes: dynamics and mechanisms of mutation. Annu. Rev. Plant Biol. 68, 225–252 (2017).

    Article  CAS  PubMed  Google Scholar 

  135. Kazama, T. et al. Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing. Nat. Plants 5, 722–730 (2019).

    Article  CAS  PubMed  Google Scholar 

  136. Arimura, S. I. et al. Targeted gene disruption of ATP synthases 6-1 and 6-2 in the mitochondrial genome of Arabidopsis thaliana by mitoTALENs. Plant J. 104, 1459–1471 (2020).

    Article  CAS  PubMed  Google Scholar 

  137. Kang, B. et al. Chloroplast and mitochondrial DNA editing in plants. Nat. Plants 7, 899–905 (2021).

    Article  PubMed  PubMed Central  Google Scholar 

  138. Bock, R. Engineering plastid genomes: methods, tools, and applications in basic research and biotechnology. Annu. Rev. Plant Biol. 66, 211–241 (2015).

    Article  CAS  PubMed  Google Scholar 

  139. Nakazato, I. et al. Targeted base editing in the plastid genome of Arabidopsis thaliana. Nat. Plants 7, 906–913 (2021).

    Article  PubMed  PubMed Central  Google Scholar 

  140. Li, R. et al. High-efficiency plastome base-editing in rice with TAL cytosine deaminase. Mol. Plant 184, 107229 (2020).

    Google Scholar 

  141. Cox, D. B. T. et al. RNA editing with CRISPR-Cas13. Science 358, 1019–1027 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  142. Liu, Y. et al. REPAIRx, a specific yet highly efficient programmable A > I RNA base editor. EMBO J. 39, e104748 (2020).

    CAS  PubMed  PubMed Central  Google Scholar 

  143. Abudayyeh, O. O. et al. A cytosine deaminase for programmable single-base RNA editing. Science 365, 382–386 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  144. Takenaka, M., Zehrmann, A., Verbitskiy, D., Härtel, B. & Brennicke, A. RNA editing in plants and its evolution. Annu. Rev. Genet. 47, 335–352 (2013).

    Article  CAS  PubMed  Google Scholar 

  145. Anzalone, A. V. et al. Search-and-replace genome editing without double-strand breaks or donor DNA. Nature 576, 149–157 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  146. Liu, Y., Kao, H. I. & Bambara, R. A. Flap endonuclease 1: a central component of DNA metabolism. Annu. Rev. Biochem. 73, 589–615 (2004).

    Article  CAS  PubMed  Google Scholar 

  147. Keijzers, G., Bohr, V. A. & Rasmussen, L. J. Human exonuclease 1 (EXO1) activity characterization and its function on flap structures. Biosci. Rep. 35, e00206 (2015).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  148. Tang, X. et al. Plant prime editors enable precise gene editing in rice cells. Mol. Plant 13, 667–670 (2020).

    Article  CAS  PubMed  Google Scholar 

  149. Lin, Q. et al. Prime genome editing in rice and wheat. Nat. Biotechnol. 38, 582–585 (2020).

    Article  CAS  PubMed  Google Scholar 

  150. Xu, R. et al. Development of plant prime-editing systems for precise genome editing. Plant Commun. 1, 100043 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  151. Butt, H. et al. Engineering herbicide resistance via prime editing in rice. Plant Biotechnol. J. 18, 2370–2372 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  152. Li, H., Li, J., Chen, J., Yan, L. & Xia, L. Precise modifications of both exogenous and endogenous genes in rice by prime editing. Mol. Plant 13, 671–674 (2020).

    Article  CAS  PubMed  Google Scholar 

  153. Xu, W. et al. Versatile nucleotides substitution in plant using an improved prime editing system. Mol. Plant 13, 675–678 (2020).

    Article  CAS  PubMed  Google Scholar 

  154. Wang, L. et al. Spelling changes and fluorescent tagging with prime editing vectors for plants. Front. Genome Ed. 3, 617553 (2021).

    Article  PubMed  PubMed Central  Google Scholar 

  155. Xu, R., Liu, X., Li, J., Qin, R. & Wei, P. Identification of herbicide resistance OsACC1 mutations via in planta prime editing-library screening in rice. Nat. Plants 7, 888–892 (2021).

    Article  CAS  PubMed  Google Scholar 

  156. Jiang, Y. Y. et al. Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize. Genome Biol. 21, 257 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  157. Veillet, F. et al. Prime editing is achievable in the tetraploid potato, but needs improvement. Preprint at bioRxiv https://doi.org/10.1101/2020.06.18.159111 (2020).

  158. Lu, Y. et al. Precise genome modification in tomato using an improved prime editing system. Plant Biotechnol. J. 19, 415–417 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  159. Lin, Q. et al. High-efficiency prime editing with optimized, paired pegRNAs in plants. Nat. Biotechnol. 39, 923–927 (2021).

    Article  CAS  PubMed  Google Scholar 

  160. Jin, S. et al. Genome-wide specificity of prime editors in plants. Nat. Biotechnol. https://doi.org/10.1038/s41587-021-00891-x (2021).

  161. Wang, J., Zhang, X., Cheng, L. & Luo, Y. An overview and metanalysis of machine and deep learning-based CRISPR gRNA design tools. RNA Biol. 17, 13–22 (2020).

    Article  CAS  PubMed  Google Scholar 

  162. Hwang, G. H. et al. Web-based design and analysis tools for CRISPR base editing. BMC Bioinform. 19, 542 (2018).

    Article  CAS  Google Scholar 

  163. Haopeng Yu, Wu,Z., Chen, X., Ji, Q. & Tao, S. CRISPR-CBEI: a designing and analyzing tool kit for cytosine. mSystems 5, e00350-20 (2020).

    PubMed  PubMed Central  Google Scholar 

  164. Dandage, R., Després, P. C., Yachie, N. & Landry, C. R. Beditor: a computational workflow for designing libraries of guide RNAs for CRISPR-mediated base editing. Genetics 212, 377–385 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  165. Arbab, M. et al. Determinants of base editing outcomes from target library analysis and machine learning. Cell 182, 463–480 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  166. Song, M. et al. Sequence-specific prediction of the efficiencies of adenine and cytosine base editors. Nat. Biotechnol. 38, 1037–1043 (2020).

    Article  CAS  PubMed  Google Scholar 

  167. Rabinowitz, R., Abadi, S., Almog, S. & Offen, D. Prediction of synonymous corrections by the BE-FF computational tool expands the targeting scope of base editing. Nucleic Acids Res. 48, W340–W347 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  168. Minkenberg, B., Zhang, J., Xie, K. & Yang, Y. CRISPR-PLANT v2: an online resource for highly specific guide RNA spacers based on improved off-target analysis. Plant Biotechnol. J. 17, 5–8 (2019).

    Article  PubMed  Google Scholar 

  169. Liu, H. et al. CRISPR-P 2.0: an improved CRISPR-Cas9 tool for genome editing in plants. Mol. Plant 10, 530–532 (2017).

    Article  CAS  PubMed  Google Scholar 

  170. Clement, K. et al. CRISPResso2 provides accurate and rapid genome editing sequence analysis. Nat. Biotechnol. 37, 224–226 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  171. Kluesner, M. G. et al. EditR: a method to quantify base editing from Sanger sequencing. CRISPR J. 1, 239–250 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  172. Xu, L., Liu, Y. & Han, R. BEAT: a Python program to quantify base editing from Sanger sequencing. CRISPR J. 2, 223–229 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  173. Chow, R. D., Chen, J. S., Shen, J. & Chen, S. A web tool for the design of prime-editing guide RNAs. Nat. Biomed. Eng. 5, 190–194 (2021).

    Article  CAS  PubMed  Google Scholar 

  174. Hsu, J. Y. et al. PrimeDesign software for rapid and simplified design of prime editing guide RNAs. Nat. Commun. 12, 8–13 (2021).

    Article  CAS  Google Scholar 

  175. Morris, J., Rahman, J., Guo, X. & Sanjana, N. Automated design of CRISPR prime editors for thousands of human pathogenic variants. Preprint at bioRxiv https://doi.org/10.1101/2020.05.07.083444 (2020).

  176. Standage-Beier, K., Tekel, S. J., Brafman, D. A. & Wang, X. Prime editing guide RNA design automation using PINE-CONE. ACS Synth. Biol. 10, 422–427 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  177. Bhagwat, A. M. et al. Multicrispr: gRNA design for prime editing and parallel targeting of thousands of targets. Life Sci. Alliance 3, e202000757 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  178. Gionfriddo, M., De Gara, L. & Loreto, F. Directed evolution of plant processes: towards a green (r)evolution? Trends Plant Sci. 24, 999–1007 (2019).

    Article  CAS  PubMed  Google Scholar 

  179. Butt, H. et al. CRISPR directed evolution of the spliceosome for resistance to splicing inhibitors. Genome Biol. 20, 73 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  180. Zhang, Y. & Qi, Y. CRISPR enables directed evolution in plants. Genome Biol. 20, 83 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  181. Ma, Y. et al. Targeted AID-mediated mutagenesis (TAM) enables efficient genomic diversification in mammalian cells. Nat. Methods 12, 1029–1035 (2016).

    Article  CAS  Google Scholar 

  182. Hess, G. T. et al. Directed evolution using dCas9-targeted somatic hypermutation in mammalian cells. Nat. Methods 12, 1036–1042 (2016).

    Article  CAS  Google Scholar 

  183. Kuang, Y. et al. Base-editing-mediated artificial evolution of OsALS1 in planta to develop novel herbicide-tolerant rice germplasms. Mol. Plant 13, 565–572 (2020).

    Article  CAS  PubMed  Google Scholar 

  184. Rodríguez-Leal, D., Lemmon, Z. H., Man, J., Bartlett, M. E. & Lippman, Z. B. Engineering quantitative trait variation for crop improvement by genome editing. Cell 171, 470–480 (2017).

    Article  PubMed  CAS  Google Scholar 

  185. Wang, X. et al. Dissecting cis-regulatory control of quantitative trait variation in a plant stem cell circuit. Nat. Plants 7, 419–427 (2021).

    Article  CAS  PubMed  Google Scholar 

  186. Zeng, D. et al. Quantitative regulation of Waxy expression by CRISPR/Cas9-based promoter and 5′UTR-intron editing improves grain quality in rice. Plant Biotechnol. J. 18, 2385–2387 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  187. Liu, L. et al. Enhancing grain-yield-related traits by CRISPR–Cas9 promoter editing of maize CLE genes. Nat. Plants 7, 287–294 (2021).

    Article  CAS  PubMed  Google Scholar 

  188. Meng, F. et al. Genomic editing of intronic enhancers unveils their role in fine-tuning tissue-specific gene expression in Arabidopsis thaliana. Plant Cell 33, 1997–2014 (2021).

    Article  PubMed  PubMed Central  Google Scholar 

  189. Zhang, H. et al. Genome editing of upstream open reading frames enables translational control in plants. Nat. Biotechnol. 36, 894–900 (2018).

    Article  CAS  PubMed  Google Scholar 

  190. Komatsu, A., Ohtake, M., Shimatani, Z. & Nishida, K. Production of herbicide-sensitive strain to prevent volunteer rice infestation using a CRISPR-Cas9 cytidine deaminase fusion. Front. Plant Sci. 11, 925 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  191. Hu, L. et al. Precision genome engineering through cytidine base editing in rapeseed (Brassica napus. L). Front. Genome Ed. 2, 605768 (2020).

    Article  PubMed  PubMed Central  Google Scholar 

  192. Wang, X. et al. Efficient gene silencing by adenine base editor-mediated start codon mutation. Mol. Ther. 28, 431–440 (2020).

    Article  CAS  PubMed  Google Scholar 

  193. Gapinske, M. et al. CRISPR-SKIP: programmable gene splicing with single base editors. Genome Biol. 19, 107 (2018).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  194. Maori, E., Galanty, Y., Pignocchi, C., GARCIA, A. C. & Meir, O. Modifying the specificity of plant non-coding RNA molecules for silencing gene expression. US patent 16/648,748 (2020).

  195. Shimatani, Z. et al. Herbicide tolerance-assisted multiplex targeted nucleotide substitution in rice. Data Brief 20, 1325–1331 (2018).

    Article  PubMed  PubMed Central  Google Scholar 

  196. Dong, H. et al. Generation of imidazolinone herbicide resistant trait in Arabidopsis. PLoS ONE 15, e0233503 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  197. Li, Y. et al. Precise base editing of non-allelic acetolactate synthase genes confers sulfonylurea herbicide resistance in maize. Crop J. 8, 449–456 (2020).

    Article  Google Scholar 

  198. Liu, X. et al. A CRISPR-Cas9-mediated domain-specific base-editing screen enables functional assessment of ACCase variants in rice. Plant Biotechnol. J. 18, 1845–1847 (2020).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  199. Liu, L. et al. Developing a novel artificial rice germplasm for dinitroaniline herbicide resistance by base editing of OsTubA2. Plant Biotechnol. J. 19, 5–7 (2021).

    Article  PubMed  CAS  Google Scholar 

  200. Hu, Z. et al. Genome editing-based engineering of cesa3 dual cellulose-inhibitor-resistant plants. Plant Physiol. 180, 827–836 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  201. Xu, Y. et al. Fine-tuning the amylose content of rice by precise base editing of the Wx gene. Plant Biotechnol. J. 19, 11–13 (2021).

    Article  CAS  PubMed  Google Scholar 

  202. Zsögön, A. et al. De novo domestication of wild tomato using genome editing. Nat. Biotechnol. 36, 1211–1216 (2018).

    Article  CAS  Google Scholar 

  203. Lemmon, Z. H. et al. Rapid improvement of domestication traits in an orphan crop by genome editing. Nat. Plants 4, 766–770 (2018).

    Article  CAS  PubMed  Google Scholar 

  204. Li, T. et al. Domestication of wild tomato is accelerated by genome editing. Nat. Biotechnol. 36, 1160–1163 (2018).

    Article  CAS  Google Scholar 

  205. Yu, H. et al. A route to de novo domestication of wild allotetraploid rice. Cell 184, 1156–1170 (2021).

    Article  CAS  PubMed  Google Scholar 

  206. Van Schie, C. C. N. & Takken, F. L. W. Susceptibility genes 101: how to be a good host. Annu. Rev. Phytopathol. 52, 551–581 (2014).

    Article  PubMed  CAS  Google Scholar 

  207. Mushtaq, M. et al. Tweaking genome-editing approaches for virus interference in crop plants. Plant Physiol. Biochem. 147, 242–250 (2020).

    Article  CAS  PubMed  Google Scholar 

  208. Langner, T., Kamoun, S. & Belhaj, K. CRISPR crops: plant genome editing toward disease resistance. Annu. Rev. Phytopathol. 56, 479–512 (2018).

    Article  CAS  PubMed  Google Scholar 

  209. Zhang, Y., Malzahn, A. A., Sretenovic, S. & Qi, Y. The emerging and uncultivated potential of CRISPR technology in plant science. Nat. Plants 5, 778–794 (2019).

    Article  PubMed  Google Scholar 

  210. Gao, C. Genome engineering for crop improvement and future agriculture. Cell 184, 1621–1635 (2021).

    Article  CAS  PubMed  Google Scholar 

  211. Nasti, R. A. & Voytas, D. F. Attaining the promise of plant gene editing at scale. Proc. Natl Acad. Sci. USA 118, e2004846117 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  212. Qin, R. et al. Increasing fidelity and efficiency by modifying cytidine base-editing systems in rice. Crop J. 8, 396–402 (2020).

    Article  Google Scholar 

  213. Siegner, S. M., Karasu, M. E., Schröder, M. S., Kontarakis, Z. & Corn, J. E. PnB Designer: a web application to design prime and base editor guide RNAs for animals and plants. BMC Bioinform. 22, 101 (2021).

    Article  CAS  Google Scholar 

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Acknowledgements

Owing to space limitations, we could not cite all of the related literature and we apologize to the authors whose work was not cited in this Review. This work was supported by the National Science Foundation Plant Genome Research Program grants (award nos IOS-1758745 and IOS-2029889), the US Department of Agriculture Biotechnology Risk Assessment Grant Program competitive grants (award nos 2018-33522-28789 and 2020-33522-32274), Emergency Citrus Disease Research and Extension Program (award no. 2020-70029-33161), Agriculture and Food Research Initiative Agricultural Innovations Through Gene Editing Program (award no. 2021-67013-34554), Foundation for Food & Agriculture Research grant (award no. 593603), Maryland Innovation Initiative Funding (award no. 1120-012_2) and Syngenta to Y.Q. K.A.M. acknowledges funding from the Indian Council of Agricultural Research (ICAR), New Delhi, in the form of the Plan Scheme—‘Incentivizing Research in Agriculture’ project and support from the Director, NRRI. S.S. is a Foundation for Food & Agriculture Research Fellow, with matching funds provided by Inari Agriculture. The content of this publication is solely the responsibility of the authors and does not necessarily represent the official views of these funding agencies. Figures were created using the BioRender tool.

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Y.Q. led the project planning. K.A.M., S.S., K.C.B. and Y.Q. wrote the manuscript. K.A.M. and S.S. prepared the figures and tables. All of the authors read and approved the final manuscript.

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Correspondence to Kutubuddin A. Molla or Yiping Qi.

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Y.Q. is a consultant for Inari Agriculture and CTC Genomics, companies that use genome editing tools in plants. The other authors declare no competing interests.

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Peer review information Nature Plants thanks Paul Hooykaas, Pengcheng Wei and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Molla, K.A., Sretenovic, S., Bansal, K.C. et al. Precise plant genome editing using base editors and prime editors. Nat. Plants 7, 1166–1187 (2021). https://doi.org/10.1038/s41477-021-00991-1

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