A multipurpose toolkit to enable advanced genome engineering in plants

Plant Cell

Volumn 29, Issue 6, 2017, Pages 1196-1217

  Čermák, Tomáš ^a   Curtin, Shaun J ^b,d   Gil Humanes, Javier ^a,e   Čegan, Radim ^c   Kono, Thomas J Y ^b   Konečná, Eva ^a   Belanto, Joseph J ^a   Starker, Colby G ^a   Mathre, Jade W ^a   Greenstein, Rebecca L ^a   Voytas, Daniel F ^a  

^a UNIVERSITY OF MINNESOTA MEDICAL SCHOOL   (United States)

^b UNIVERSITY OF MINNESOTA   (United States)

^c INSTITUTE OF MATHEMATICS   (Czech Republic)

^d AGRICULTURAL RESEARCH SERVICE   (United States)

^e Calyxt Inc   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

PLANT PROTEIN; PLANT RNA; TRANSCRIPTION ACTIVATOR LIKE EFFECTOR NUCLEASE;

CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEAT; GENETIC ENGINEERING; GENETICS; HORDEUM; PROCEDURES; TOMATO; TRANSGENIC PLANT; WHEAT;

CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS; GENETIC ENGINEERING; HORDEUM; LYCOPERSICON ESCULENTUM; PLANT PROTEINS; PLANTS, GENETICALLY MODIFIED; RNA, PLANT; TRANSCRIPTION ACTIVATOR-LIKE EFFECTOR NUCLEASES; TRITICUM;

EID: 85025685430     PISSN: 10404651     EISSN: 1532298X     Source Type: Journal    
DOI: 10.1105/tpc.16.00922     Document Type: Article

References (91)

1. Aeby, E., Ullu, E., Yepiskoposyan, H., Schimanski, B., Roditi, I., Mühlemann, O., and Schneider, A. (2010). tRNASec is transcribed by RNA polymerase II in Trypanosoma brucei but not in humans. Nucleic Acids Res. 38: 5833–5843.
2. Arimbasseri, A.G., Rijal, K., and Maraia, R.J. (2013). Transcription termination by the eukaryotic RNA polymerase III. Biochim. Biophys. Acta 1829: 318–330.
3. Baltes, N.J., Gil-Humanes, J., Cermak, T., Atkins, P.A., and Voytas, D.F. (2014). DNA replicons for plant genome engineering. Plant Cell 26: 151–163.
4. Baltes, N.J., and Voytas, D.F. (2015). Enabling plant synthetic biology through genome engineering. Trends Biotechnol. 33: 120-131.
5. Brooks, C., Nekrasov, V., Lippman, Z.B., and Van Eck, J. (2014). Efficient gene editing in tomato in the first generation using the clustered regularly interspaced short palindromic repeats/CRISPRassociated9 system. Plant Physiol. 166: 1292–1297.
6. Canver, M.C., Bauer, D.E., Dass, A., Yien, Y.Y., Chung, J., Masuda, T., Maeda, T., Paw, B.H., and Orkin, S.H. (2014). Characterization of genomic deletion efficiency mediated by clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 nuclease system in mammalian cells. J. Biol. Chem. 289: 21312–21324.
7. Cěrmák, T., Baltes, N.J., Cěgan, R., Zhang, Y., and Voytas, D.F. (2015). High-frequency, precise modification of the tomato genome. Genome Biol. 16: 232.
8. Cermak, T., Doyle, E.L., Christian, M., Wang, L., Zhang, Y., Schmidt, C., Baller, J.A., Somia, N.V., Bogdanove, A.J., and Voytas, D.F. (2011). Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting. Nucleic Acids Res. 39: e82.
9. Certo, M.T., et al. (2012). Coupling endonucleases with DNA end-processing enzymes to drive gene disruption. Nat. Methods 9: 973–975.
10. Clasen, B.M., et al. (2016). Improving cold storage and processing traits in potato through targeted gene knockout. Plant Biotechnol. J. 14: 169–176.
11. Cong, L., Ran, F.A., Cox, D., Lin, S., Barretto, R., Habib, N., Hsu, P.D., Wu, X., Jiang, W., Marraffini, L.A., and Zhang, F. (2013). Multiplex genome engineering using CRISPR/Cas systems. Science 339: 819–823.
12. Cosson, V., Durand, P., d’Erfurth, I., Kondorosi, A., and Ratet, P. (2006). Medicago truncatula transformation using leaf explants. Methods Mol. Biol. 343: 115–127.
13. Curtin, S.J., Tiffin, P., Guhlin, J., Trujillo, D.I., Burghart, L.T., Atkins, P., Baltes, N.J., Denny, R., Voytas, D.F., Stupar, R.M., and Young, N.D. (2017). Validating Genome-Wide Association candidates through quantitative variation in nodulation. Plant Physiol. 173: 921–931.
14. Curtis, M.D., and Grossniklaus, U. (2003). A gateway cloning vector set for high-throughput functional analysis of genes in planta. Plant Physiol. 133: 462–469.
15. Doyon, Y., Vo, T.D., Mendel, M.C., Greenberg, S.G., Wang, J., Xia, D.F., Miller, J.C., Urnov, F.D., Gregory, P.D., and Holmes, M.C. (2011). Enhancing zinc-finger-nuclease activity with improved obligate heterodimeric architectures. Nat. Methods 8: 74–79.
16. Engler, C., Gruetzner, R., Kandzia, R., and Marillonnet, S. (2009). Golden gate shuffling: a one-pot DNA shuffling method based on type IIs restriction enzymes. PLoS One 4: e5553.
17. Engler, C., Kandzia, R., and Marillonnet, S. (2008). A one pot, one step, precision cloning method with high throughput capability. PLoS One 3: e3647.
18. Expósito-Rodríguez, M., Borges, A.A., Borges-Pérez, A., and Pérez, J.A. (2008). Selection of internal control genes for quantitative real-time RT-PCR studies during tomato development process. BMC Plant Biol. 8: 131.
19. Fauser, F., Roth, N., Pacher, M., Ilg, G., Sánchez-Fernández, R., Biesgen, C., and Puchta, H. (2012). In planta gene targeting. Proc. Natl. Acad. Sci. USA 109: 7535–7540.
20. Fauser, F., Schiml, S., and Puchta, H. (2014). Both CRISPR/Casbased nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana. Plant J. 79: 348–359.
21. Forner, J., Pfeiffer, A., Langenecker, T., Manavella, P.A., and Lohmann, J.U. (2015). Germline-transmitted genome editing in Arabidopsis thaliana Using TAL-effector-nucleases. PLoS One 10: e0121056. Erratum. PLoS One 10: e0133945.
22. Fu, Y., Sander, J.D., Reyon, D., Cascio, V.M., and Joung, J.K. (2014). Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat. Biotechnol. 32: 279–284.
23. Gao, J., Wang, G., Ma, S., Xie, X., Wu, X., Zhang, X., Wu, Y., Zhao, P., and Xia, Q. (2015). CRISPR/Cas9-mediated targeted mutagenesis in Nicotiana tabacum. Plant Mol. Biol. 87: 99–110.
24. Gao, Y., and Zhao, Y. (2014). Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing. J. Integr. Plant Biol. 56: 343–349.
25. Geisinger, J.M., Turan, S., Hernandez, S., Spector, L.P., and Calos, M.P. (2016). In vivo blunt-end cloning through CRISPR/Cas9-facilitated non-homologous end-joining. Nucleic Acids Res. 44: e76.
26. Gibson, D.G., Young, L., Chuang, R.Y., Venter, J.C., Hutchison III C.A., and Smith, H.O. (2009). Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat. Methods 6: 343–345.
27. Gil-Humanes, J., Ozuna, C.V., Marín, S., León, E., Barro, F., and Pistón, F. (2011). Genetic transformation of wheat: advances in the transformation method and applications for obtaining lines with improved bread-making quality and low toxicity in relation to celiac disease. In Genetic Transformation, M. Alvarez, ed (InTech), pp. 135–150.
28. Gil-Humanes, J., Wang, Y., Liang, Z., Shan, Q., Ozuna, C.V., Sánchez-León, S., Baltes, N.J., Starker, C., Barro, F., Gao, C., and Voytas, D.F. (2017). High-efficiency gene targeting in hexaploid wheat using DNA replicons and CRISPR/Cas9. Plant J. 89: 1251-1262.
29. Guilinger, J.P., Thompson, D.B., and Liu, D.R. (2014). Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification. Nat. Biotechnol. 32: 577–582.
30. Haseloff, J., and Gerlach, W.L. (1988). Simple RNA enzymes with new and highly specific endoribonuclease activities. Nature 334: 585–591.
31. Haun, W., et al. (2014). Improved soybean oil quality by targeted mutagenesis of the fatty acid desaturase 2 gene family. Plant Biotechnol. J. 12: 934–940.
32. Chari, R., Mali, P., Moosburner, M., and Church, G.M. (2015). Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach. Nat. Methods 12: 823–826.
33. Chavez, A., et al. (2015). Highly efficient Cas9-mediated transcriptional programming. Nat. Methods 12: 326–328.
34. Chen, B., Gilbert, L.A., Cimini, B.A., Schnitzbauer, J., Zhang, W., Li, G.W., Park, J., Blackburn, E.H., Weissman, J.S., Qi, L.S., and Huang, B. (2013). Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system. Cell 155: 1479–1491.
35. Cho, S.W., Kim, S., Kim, Y., Kweon, J., Kim, H.S., Bae, S., and Kim, J.S. (2014). Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases. Genome Res. 24: 132-141.
36. Christensen, A.H., and Quail, P.H. (1996). Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants. Transgenic Res. 5: 213–218.
37. Christian, M., Qi, Y., Zhang, Y., and Voytas, D.F. (2013). Targeted mutagenesis of Arabidopsis thaliana using engineered TAL effector nucleases. G3 (Bethesda) 3: 1697–1705.
38. Jinek, M., Chylinski, K., Fonfara, I., Hauer, M., Doudna, J.A., and Charpentier, E. (2012). A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337: 816-821.
39. Kim, H., Kim, S.T., Ryu, J., Choi, M.K., Kweon, J., Kang, B.C., Ahn, H.M., Bae, S., Kim, J., Kim, J.S., and Kim, S.G. (2016). A simple, flexible and high-throughput cloning system for plant genome editing via CRISPR-Cas system. J. Integr. Plant Biol. 58: 705–712.
40. Kungulovski, G., and Jeltsch, A. (2016). Epigenome Editing: State of the Art, Concepts, and Perspectives. Trends Genet. 32: 101–113.
41. Kwon, Y.I., Abe, K., Osakabe, K., Endo, M., Nishizawa-Yokoi, A., Saika, H., Shimada, H., and Toki, S. (2012). Overexpression of OsRecQl4 and/or OsExo1 enhances DSB-induced homologous recombination in rice. Plant Cell Physiol. 53: 2142–2152.
42. La Russa, M.F., and Qi, L.S. (2015). The new state of the art: Cas9 for gene activation and repression. Mol. Cell. Biol. 35: 3800-3809.
43. Li, J., et al. (2016). Multiplexed, targeted gene editing in Nicotiana benthamiana for glyco-engineering and monoclonal antibody production. Plant Biotechnol. J. 14: 533–542.
44. Li, T., Liu, B., Spalding, M.H., Weeks, D.P., and Yang, B. (2012). High-efficiency TALEN-based gene editing produces disease-resistant rice. Nat. Biotechnol. 30: 390–392.
45. Li, Y., et al. (2015). A versatile reporter system for CRISPR-mediated chromosomal rearrangements. Genome Biol. 16: 111.
46. Liang, G., Zhang, H., Lou, D., and Yu, D. (2016). Selection of highly efficient sgRNAs for CRISPR/Cas9-based plant genome editing. Sci. Rep. 6: 21451.
47. Liang, Z., Zhang, K., Chen, K., and Gao, C. (2014). Targeted mutagenesis in Zea mays using TALENs and the CRISPR/Cas system. J. Genet. Genomics 41: 63–68.
48. Liu, N., Wu, S., Van Houten, J., Wang, Y., Ding, B., Fei, Z., Clarke, T.H., Reed, J.W., and van der Knaap, E. (2014). Down-regulation of AUXIN RESPONSE FACTORS 6 and 8 by microRNA 167 leads to floral development defects and female sterility in tomato. J. Exp. Bot. 65: 2507–2520.
49. Liu, W., Rudis, M.R., Peng, Y., Mazarei, M., Millwood, R.J., Yang, J.P., Xu, W., Chesnut, J.D., and Stewart, C.N., Jr. (2014). Synthetic TAL effectors for targeted enhancement of transgene expression in plants. Plant Biotechnol. J. 12: 436–446.
50. Lowder, L.G., Zhang, D., Baltes, N.J., Paul J.W., Tang, X., Zheng, X., Voytas, D.F., Hsieh, T.-F., Zhang, Y., and Qi, Y. (2015). A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation. Plant Physiol. 169: 971–985.
51. Luo, S., Li, J., Stoddard, T.J., Baltes, N.J., Demorest, Z.L., Clasen, B.M., Coffman, A., Retterath, A., Mathis, L., Voytas, D.F., and Zhang, F. (2015). Non-transgenic plant genome editing using purified sequence-specific nucleases. Mol. Plant 8: 1425–1427.
52. Ma, X., et al. (2015). A robustCRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicotplants. Mol. Plant 8: 1474–1484.
53. Maiti, I.B., Gowda, S., Kiernan, J., Ghosh, S.K., and Shepherd, R.J. (1997). Promoter/leader deletion analysis and plant expression vectors with the figwort mosaic virus (FMV) full length transcript (FLt) promoter containing single or double enhancer domains. Transgenic Res. 6: 143–156.
54. Mali, P., Aach, J., Stranges, P.B., Esvelt, K.M., Moosburner, M., Kosuri, S., Yang, L., and Church, G.M. (2013). CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nat. Biotechnol. 31: 833–838.
55. Mann, D.G., et al. (2011). Switchgrass (Panicum virgatum L.) polyubiquitin gene (PvUbi1 and PvUbi2) promoters for use in plant transformation. BMC Biotechnol. 11: 74.
56. Mann, D.G.J., Lafayette, P.R., Abercrombie, L.L., King, Z.R., Mazarei, M., Halter, M.C., Poovaiah, C.R., Baxter, H., Shen, H., Dixon, R.A., Parrott, W.A., and Neal Stewart, C., Jr. (2012). Gateway-compatible vectors for high-throughput gene functional analysis in switchgrass (Panicum virgatum L.) and other monocot species. Plant Biotechnol. J. 10: 226–236.
57. McElroy, D., Zhang, W., Cao, J., and Wu, R. (1990). Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2: 163–171.
58. Mikami, M., Toki, S., and Endo, M. (2016). Precision targeted mutagenesis via Cas9 paired nickases in rice. Plant Cell Physiol. 57: 1058–1068.
59. Miller, J.C., et al. (2011). A TALE nuclease architecture for efficient genome editing. Nat. Biotechnol. 29: 143–148.
60. Nagaya, S., Kawamura, K., Shinmyo, A., and Kato, K. (2010). The HSP terminator of Arabidopsis thaliana increases gene expression in plant cells. Plant Cell Physiol. 51: 328–332.
61. Nissim, L., Perli, S.D., Fridkin, A., Perez-Pinera, P., and Lu, T.K. (2014). Multiplexed and programmable regulation of gene networks with an integrated RNA and CRISPR/Cas toolkit in human cells. Mol. Cell 54: 698–710.
62. Norris, S.R., Meyer, S.E., and Callis, J. (1993). The intron of Arabidopsis thaliana polyubiquitin genes is conserved in location and is a quantitative determinant of chimeric gene expression. Plant Mol. Biol. 21: 895–906.
63. Ordon, J., Gantner, J., Kemna, J., Schwalgun, L., Reschke, M., Streubel, J., Boch, J., and Stuttmann, J. (2017). Generation of chromosomal deletions in dicotyledonous plants employing a userfriendly genome editing toolkit. Plant J. 89: 155–168.
64. Park, B.S., Seo, J.S., and Chua, N.H. (2014). NITROGEN LIMITATION ADAPTATION recruits PHOSPHATE2 to target the phosphate transporter PT2 for degradation during the regulation of Arabidopsis phosphate homeostasis. Plant Cell 26: 454–464.
65. Piatek, A., Ali, Z., Baazim, H., Li, L., Abulfaraj, A., Al-Shareef, S., Aouida, M., and Mahfouz, M.M. (2015). RNA-guided transcriptional regulation in planta via synthetic dCas9-based transcription factors. Plant Biotechnol. J. 13: 578–589.
66. 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., and Zhang, F. (2013). Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell 154: 1380–1389.
67. Sahoo, D.K., Dey, N., and Maiti, I.B. (2014). pSiM24 is a novel versatile gene expression vector for transient assays as well as stable expression of foreign genes in plants. PLoS One 9: e98988.
68. Shan, Q., Wang, Y., Li, J., Zhang, Y., Chen, K., Liang, Z., Zhang, K., Liu, J., Xi, J.J., Qiu, J.-L., and Gao, C. (2013). Targeted genome modification of crop plants using a CRISPR-Cas system. Nat. Biotechnol. 31: 686–688.
69. Shen, B., Zhang, W., Zhang, J., Zhou, J., Wang, J., Chen, L., Wang, L., Hodgkins, A., Iyer, V., Huang, X., and Skarnes, W.C. (2014). Efficient genome modification by CRISPR-Cas9 nickase with minimal off-target effects. Nat. Methods 11: 399–402.
70. Shimatani, Z., et al. (2017). Targeted base editing in rice and tomato using a CRISPR-Cas9 cytidine deaminase fusion. Nat. Biotechnol. 35: 441–443.
71. Schiml, S., Fauser, F., and Puchta, H. (2014). 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.
72. Schneider, C.A., Rasband, W.S., and Eliceiri, K.W. (2012). NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9: 671-675.
73. Stavolone, L., Kononova, M., Pauli, S., Ragozzino, A., de Haan, P., Milligan, S., Lawton, K., and Hohn, T. (2003). Cestrum yellow leaf curling virus (CmYLCV) promoter: a new strong constitutive promoter for heterologous gene expression in a wide variety of crops. Plant Mol. Biol. 53: 663–673.
74. Sugano, S.S., Shirakawa, M., Takagi, J., Matsuda, Y., Shimada, T., Hara-Nishimura, I., and Kohchi, T. (2014). CRISPR/Cas9-mediated targeted mutagenesis in the liverwort Marchantia polymorpha L. Plant Cell Physiol. 55: 475–481.
75. Sugio, T., Satoh, J., Matsuura, H., Shinmyo, A., and Kato, K. (2008). The 59-untranslated region of the Oryza sativa alcohol dehydrogenase gene functions as a translational enhancer in monocotyledonous plant cells. J. Biosci. Bioeng. 105: 300–302.
76. Tang, X., Zheng, X., Qi, Y., Zhang, D., Cheng, Y., Tang, A., Voytas, D.F., and Zhang, Y. (2016). A single transcript CRISPR-Cas9 system for efficient genome editing in plants. Mol. Plant 9: 1088–1091.
77. Thole, V., Worland, B., Snape, J.W., and Vain, P. (2007). The pCLEAN dual binary vector system for Agrobacterium-mediated plant transformation. Plant Physiol. 145: 1211–1219.
78. Tsai, S.Q., Wyvekens, N., Khayter, C., Foden, J.A., Thapar, V., Reyon, D., Goodwin, M.J., Aryee, M.J., and Joung, J.K. (2014). Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nat. Biotechnol. 32: 569–576.
79. Vazquez-Vilar, M., Bernabé-Orts, J.M., Fernandez-Del-Carmen, A., Ziarsolo, P., Blanca, J., Granell, A., and Orzaez, D. (2016). A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. Plant Methods 12: 10.
80. Wang, Y., Cheng, X., Shan, Q., Zhang, Y., Liu, J., Gao, C., and Qiu, J.L. (2014). Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew. Nat. Biotechnol. 32: 947–951.
81. Wang, Z.P., Xing, H.L., Dong, L., Zhang, H.Y., Han, C.Y., Wang, X.C., and Chen, Q.J. (2015). Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently generates homozygous mutants for multiple target genes in Arabidopsis in a single generation. Genome Biol. 16: 144.
82. Wright, D.A., Townsend, J.A., Winfrey, R.J., Jr., Irwin, P.A., Rajagopal, J., Lonosky, P.M., Hall, B.D., Jondle, M.D., and Voytas, D.F. (2005). High-frequency homologous recombination in plants mediated by zinc-finger nucleases. Plant J. 44: 693–705.
83. Xie, K., Minkenberg, B., and Yang, Y. (2015). Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proc. Natl. Acad. Sci. USA 112: 3570–3575.
84. Xing, H.L., Dong, L., Wang, Z.P., Zhang, H.Y., Han, C.Y., Liu, B., Wang, X.C., and Chen, Q.J. (2014). A CRISPR/Cas9 toolkit for multiplex genome editing in plants. BMC Plant Biol. 14: 327.
85. Xu, R.F., Li, H., Qin, R.Y., Li, J., Qiu, C.H., Yang, Y.C., Ma, H., Li, L., Wei, P.C., and Yang, J.B. (2015). Generation of inheritable and “transgene clean” targeted genome-modified rice in later generations using the CRISPR/Cas9 system. Sci. Rep. 5: 11491.
86. Yan, L., Wei, S., Wu, Y., Hu, R., Li, H., Yang, W., and Xie, Q. (2015). High efficiency genome editing in Arabidopsis using Yao promoterdriven CRISPR/Cas9 system. Mol. Plant 8: 1820–1823.
87. Zhang, F., Maeder, M.L., Unger-Wallace, E., Hoshaw, J.P., Reyon, D., Christian, M., Li, X., Pierick, C.J., Dobbs, D., Peterson, T., Joung, J.K., and Voytas, D.F. (2010). High frequency targeted mutagenesis in Arabidopsis thaliana using zinc finger nucleases. Proc. Natl. Acad. Sci. USA 107: 12028–12033.
88. Zhang, H., Zhang, J., Wei, P., Zhang, B., Gou, F., Feng, Z., Mao, Y., Yang, L., Zhang, H., Xu, N., and Zhu, J.K. (2014). The CRISPR/ Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnol. J. 12: 797–807.
89. Zhang, Y., Zhang, F., Li, X., Baller, J.A., Qi, Y., Starker, C.G., Bogdanove, A.J., and Voytas, D.F. (2013). Transcription activatorlike effector nucleases enable efficient plant genome engineering. Plant Physiol. 161: 20–27.
90. Zhang, Z., Mao, Y., Ha, S., Liu, W., Botella, J.R., and Zhu, J.K. (2016). A multiplex CRISPR/Cas9 platform for fast and efficient editing of multiple genes in Arabidopsis. Plant Cell Rep. 35: 1519-1533.
91. Zong, Y., Wang, Y., Li, C., Zhang, R., Chen, K., Ran, Y., Qiu, J.L., Wang, D., and Gao, C. (2017). Precise base editing in rice, wheat and maize with a Cas9-cytidine deaminase fusion. Nat. Biotechnol. 35: 438–440.