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Vol. 64 No. 14: Issue 14

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CRISPR/Cas9; A robust technology for producing genetically engineered plants

https://doi.org/10.14715/cmb/2018.64.14.6
Rimsha Farooq
Department of Bioinformatics and Biotechnology, Government College University Faisalabad, Pakistan
Khadim Hussain
Department of Bioinformatics and Biotechnology, Government College University Faisalabad, Pakistan
Shahid Nazir
Agriculture biotechnology research institute, Ayub Agriculture Research Institute (AARI), Faisalabad, Pakistan
Muhammad Rizwan Javed
Department of Bioinformatics and Biotechnology, Government College University Faisalabad, Pakistan
Nazish Masood
Department of Bioinformatics and Biotechnology, Government College University Faisalabad, Pakistan

Corresponding Author(s) : Khadim Hussain

hussaink@gcuf.edu.pk

Cellular and Molecular Biology, Vol. 64 No. 14: Issue 14

Abstract

CRISPR/Cas9 is a technology evolved from modified type II immune system of bacteria and archaea. Exploitation of this bacterial immune system in all eukaryotes including plants may lead to site-specific targeted genome engineering. Genome engineering is objectively utilized to express/silence a trait harbouring gene in the plant genome. In this review, different genetic engineering techniques including classical breeding, RNAi and genetic transformation and synthetic sequence-specific nucleases (zinc finger nucleases; ZFNs and transcription activator-like effector nuclease; TALENs) techniques have been described and compared with advanced genome editing technique CRISPR/Cas9, on the basis of their merits and drawbacks. This revolutionary genome engineering technology has edge over all other approaches because of its simplicity, stability, specificity of the target and multiple genes can be engineered at a time. CRISPR/Cas9 requires only Cas9 endonuclease and single guide RNA, which are directly delivered into plant cells via either vector-mediated stable transformation or transient delivery of ribonucleoproteins (RNPs) and generate double-strand breaks (DSBs) at target site. These DSBs are further repaired by cell endogenous repairing pathways via HDR or NHEJ. The major advantage of CRISPR/Cas9 system is that engineered plants are considered Non-GM; can be achieved using in vitro expressed RNPs transient delivery. Different variants of Cas9 genes cloned in different plasmid vectors can be used to achieve different objectives of genome editing including double-stranded DNA break, single-stranded break, activate/repress the gene expression. Fusion of Cas9 with fluorescent protein can lead to visualize the expression of the CRISPR/Cas9 system. The applications of this technology in plant genome editing to improve different plant traits are comprehensively described.

Keywords

Plant genome editing CRISPR/Cas9 ZFNs TALENs RNPs.
Farooq, R., Hussain, K., Nazir, S., Javed, M. R., & Masood, N. (2018). CRISPR/Cas9; A robust technology for producing genetically engineered plants. Cellular and Molecular Biology, 64(14), 31–38. https://doi.org/10.14715/cmb/2018.64.14.6
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References
  1. M. Endo, M. Mikami and S. Toki Biallelic gene targeting in rice Plant physiology 170 (2016) 667-677.
  2. T. Cermak, E. L. Doyle, M. Christian, L. Wang, Y. Zhang, C. Schmidt, J. A. Baller, N. V. Somia, A. J. Bogdanove and D. F. Voytas Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting Nucleic Acids Res 39 e82.
  3. S. Svitashev, J. Young, C. Schwartz, H. Gao, S. C. Falco and A. M. Cigan Targeted mutagenesis, precise gene editing and site-specific gene insertion in maize using Cas9 and guide RNA Plant physiology (2015) pp. 00793.02015.
  4. Z. Li, Z.-B. Liu, A. Xing, B. P. Moon, J. P. Koellhoffer, L. Huang, R. T. Ward, E. Clifton, S. C. Falco and A. M. Cigan Cas9-guide RNA directed genome editing in soybean Plant physiology 169 (2015) 960-970.
  5. R. R. Beerli, B. Dreier and C. F. Barbas Positive and negative regulation of endogenous genes by designed transcription factors Proceedings of the National Academy of Sciences 97 (2000) 1495-1500.
  6. B. Chen, L. A. Gilbert, B. A. Cimini, J. Schnitzbauer, W. Zhang, G. W. Li, J. Park, E. H. Blackburn, J. S. Weissman, L. S. Qi and B. Huang Dynamic imaging of genomic loci in living human cells by an optimized CRISPR/Cas system Cell 155 (2013) 1479-1491.
  7. M. Luo, B. Gilbert and M. Ayliffe Applications of CRISPR/Cas9 technology for targeted mutagenesis, gene replacement and stacking of genes in higher plants Plant cell reports 35 (2016) 1439-1450.
  8. T. O. Auer, K. Duroure, A. De Cian, J.-P. Concordet and F. Del Bene Highly efficient CRISPR/Cas9-mediated knock-in in zebrafish by homology-independent DNA repair Genome Research 24 (2014) 142-153.
  9. J. Steinert, S. Schiml and H. Puchta Homology-based double-strand break-induced genome engineering in plants Plant cell reports 35 (2016) 1429-1438.
  10. F. J. Mojica, J. Garcí­a-Martí­nez and E. Soria Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements Journal of molecular evolution 60 (2005) 174-182.
  11. T. Ishii and M. Araki Consumer acceptance of food crops developed by genome editing Plant cell reports 35 (2016) 1507-1518.
  12. S. Peng, J. P. York and P. Zhang A transgenic approach for RNA interference-based genetic screening in mice Proceedings of the National Academy of Sciences of the United States of America 103 (2006) 2252-2256.
  13. L. Pena Transgenic plants: Methods and protocols, Springer Science & Business Media, 2005.
  14. D. P. Weeks, M. H. Spalding and B. Yang Use of designer nucleases for targeted gene and genome editing in plants Plant Biotechnology Journal 14 (2016) 483-495.
  15. E. Southgate, M. Davey, J. Power and R. Marchant Factors affecting the genetic engineering of plants by microprojectile bombardment Biotechnology advances 13 (1995) 631-651.
  16. N. G. Bologna and O. Voinnet The diversity, biogenesis, and activities of endogenous silencing small RNAs in Arabidopsis Annual review of plant biology 65 (2014) 473-503.
  17. D. C. Baulcombe VIGS, HIGS and FIGS: small RNA silencing in the interactions of viruses or filamentous organisms with their plant hosts Current opinion in plant biology 26 (2015) 141-146.
  18. M. Sticklen Plant genetic engineering to improve biomass characteristics for biofuels Current opinion in biotechnology 17 (2006) 315-319.
  19. Z. Zhang, Y. Mao, S. Ha, W. Liu, J. R. Botella and J.-K. Zhu A multiplex CRISPR/Cas9 platform for fast and efficient editing of multiple genes in Arabidopsis Plant Cell Reports 35 (2016) 1519-1533.
  20. N. Agrawal, P. V. Dasaradhi, A. Mohmmed, P. Malhotra, R. K. Bhatnagar and S. K. Mukherjee RNA interference: biology, mechanism, and applications Microbiol Mol Biol Rev 67 (2003) 657-685.
  21. K. T. Love, K. P. Mahon, C. G. Levins, K. A. Whitehead, W. Querbes, J. R. Dorkin, J. Qin, W. Cantley, L. L. Qin and T. Racie Lipid-like materials for low-dose, in vivo gene silencing Proceedings of the National Academy of Sciences 107 (2010) 1864-1869.
  22. E. Breese and M. Hayward The genetic basis of present breeding methods in forage crops Euphytica 21 (1972) 324-336.
  23. M. Adem APPLICATION OF PRECISE GENOME EDITING IN PLANTS SDRP Journal of Plant Science 2 (2017) 1-9.
  24. J. R. Latham, A. K. Wilson and R. A. Steinbrecher The mutational consequences of plant transformation BioMed Research International 2006 (2006) 1-7.
  25. M. Antoniou, C. Robinson and J. Fagan GMO myths and truths Earth Open Source 47 (2012).
  26. J. Van der Oost, M. M. Jore, E. R. Westra, M. Lundgren and S. J. Brouns CRISPR-based adaptive and heritable immunity in prokaryotes Trends in biochemical sciences 34 (2009) 401-407.
  27. S. Eyre, J. Bowes, D. e. Diogo, A. Lee, A. Barton, P. Martin, A. Zhernakova, E. Stahl, S. Viatte and K. McAllister High-density genetic mapping identifies new susceptibility loci for rheumatoid arthritis Nature genetics 44 (2012) 1336.
  28. I. Grissa, G. Vergnaud and C. Pourcel CRISPRFinder: a web tool to identify clustered regularly interspaced short palindromic repeats Nucleic acids research 35 (2007) W52-W57.
  29. W. D. Travis, T. E. King, E. D. Bateman, D. A. Lynch, F. d. Capron, D. Center, T. V. Colby, J. F. o. Cordier, R. M. DuBois and J. Galvin American Thoracic Society/European Respiratory Society international multidisciplinary consensus classification of the idiopathic interstitial pneumonias American Journal of Respiratory and Critical Care Medicine 165 (2002) 277-304.
  30. E. Deltcheva, K. Chylinski, C. M. Sharma, K. Gonzales, Y. Chao, Z. A. Pirzada, M. R. Eckert, J. r. Vogel and E. Charpentier CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III Nature 471 (2011) 602.
  31. M. Jinek, K. Chylinski, I. Fonfara, M. Hauer, J. A. Doudna and E. Charpentier A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity science 337 (2012) 816-821.
  32. J. E. Garneau, M. E. Dupuis, M. Villion, D. A. Romero, R. Barrangou, P. Boyaval, C. Fremaux, P. Horvath, A. H. Magadan and S. Moineau The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA Nature 468 (2010) 67-71.
  33. M. Szczepek, V. Brondani, J. Büchel, L. Serrano, D. J. Segal and T. Cathomen Structure-based redesign of the dimerization interface reduces the toxicity of zinc-finger nucleases Nature biotechnology 25 (2007) 786.
  34. N. P. Pavletich and C. O. Pabo Zinc finger-DNA recognition: crystal structure of a Zif268-DNA complex at 2.1 A Science 252 (1991) 809-817.
  35. H. A. Greisman and C. O. Pabo A general strategy for selecting high-affinity zinc finger proteins for diverse DNA target sites Science 275 (1997) 657-661.
  36. M. Isalan, A. Klug and Y. Choo A rapid, generally applicable method to engineer zinc fingers illustrated by targeting the HIV-1 promoter Nature biotechnology 19 (2001) 656.
  37. R. R. Beerli and C. F. Barbas III Engineering polydactyl zinc-finger transcription factors Nature biotechnology 20 (2002) 135.
  38. M. L. Maeder, S. Thibodeau-Beganny, A. Osiak, D. A. Wright, R. M. Anthony, M. Eichtinger, T. Jiang, J. E. Foley, R. J. Winfrey and J. A. Townsend Rapid "open-source” engineering of customized zinc-finger nucleases for highly efficient gene modification Molecular cell 31 (2008) 294-301.
  39. J. D. Sander, E. J. Dahlborg, M. J. Goodwin, L. Cade, F. Zhang, D. Cifuentes, S. J. Curtin, J. S. Blackburn, S. Thibodeau-Beganny and Y. Qi Selection-free zinc-finger-nuclease engineering by context-dependent assembly (CoDA) Nature methods 8 (2011) 67.
  40. T. Cathomen and M. Weitzman Gene repair: pointing the finger at genetic disease Gene Therapy 12 (2005) 1415-1416.
  41. J. K. Joung and J. D. Sander TALENs: a widely applicable technology for targeted genome editing Nature reviews Molecular cell biology 14 (2013) 49.
  42. D. Deng, C. Yan, X. Pan, M. Mahfouz, J. Wang, J.-K. Zhu, Y. Shi and N. Yan Structural basis for sequence-specific recognition of DNA by TAL effectors Science 335 (2012) 720-723.
  43. A. N.-S. Mak, P. Bradley, R. A. Cernadas, A. J. Bogdanove and B. L. Stoddard The crystal structure of TAL effector PthXo1 bound to its DNA target Science 335 (2012) 716-719.
  44. S. W. Cho, S. Kim, Y. Kim, J. Kweon, H. S. Kim, S. Bae and J. S. Kim Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases Genome Res 24 132-141.
  45. J. P. Guilinger, V. Pattanayak, D. Reyon, S. Q. Tsai, J. D. Sander, J. K. Joung and D. R. Liu Broad specificity profiling of TALENs results in engineered nucleases with improved DNA-cleavage specificity Nature methods 11 (2014) 429.
  46. T. Li, S. Huang, X. Zhao, D. A. Wright, S. Carpenter, M. H. Spalding, D. P. Weeks and B. Yang Modularly assembled designer TAL effector nucleases for targeted gene knockout and gene replacement in eukaryotes Nucleic acids research 39 (2011) 6315-6325.
  47. D. Reyon, S. Q. Tsai, C. Khayter, J. A. Foden, J. D. Sander and J. K. Joung FLASH assembly of TALENs for high-throughput genome editing Nature biotechnology 30 (2012) 460.
  48. D. Hockemeyer, H. Wang, S. Kiani, C. S. Lai, Q. Gao, J. P. Cassady, G. J. Cost, L. Zhang, Y. Santiago and J. C. Miller Genetic engineering of human pluripotent cells using TALE nucleases Nature biotechnology 29 (2011) 731.
  49. Y. Fu, J. A. Foden, C. Khayter, M. L. Maeder, D. Reyon, J. K. Joung and J. D. Sander High-frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells Nature biotechnology 31 (2013) 822.
  50. P. D. Hsu, D. A. Scott, J. A. Weinstein, F. A. Ran, S. Konermann, V. Agarwala, Y. Li, E. J. Fine, X. Wu and O. Shalem DNA targeting specificity of RNA-guided Cas9 nucleases Nature biotechnology 31 (2013) 827.
  51. P. Mali, J. Aach, P. B. Stranges, K. M. Esvelt, M. Moosburner, S. Kosuri, L. Yang and G. M. Church CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering Nature biotechnology 31 (2013) 833.
  52. V. Pattanayak, S. Lin, J. P. Guilinger, E. Ma, J. A. Doudna and D. R. Liu High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity Nature biotechnology 31 (2013) 839.
  53. C. Smith, A. Gore, W. Yan, L. Abalde-Atristain, Z. Li, C. He, Y. Wang, R. A. Brodsky, K. Zhang and L. Cheng Whole-genome sequencing analysis reveals high specificity of CRISPR/Cas9 and TALEN-based genome editing in human iPSCs Cell stem cell 15 (2014) 12-13.
  54. K. Suzuki, C. Yu, J. Qu, M. Li, X. Yao, T. Yuan, A. Goebl, S. Tang, R. Ren and E. Aizawa Targeted gene correction minimally impacts whole-genome mutational load in human-disease-specific induced pluripotent stem cell clones Cell stem cell 15 (2014) 31-36.
  55. B. Ellis, M. Hirsch, S. Porter, R. Samulski and M. Porteus Zinc-finger nuclease-mediated gene correction using single AAV vector transduction and enhancement by Food and Drug Administration-approved drugs Gene therapy 20 (2013) 35.
  56. M. Holkers, I. Maggio, J. Liu, J. M. Janssen, F. Miselli, C. Mussolino, A. Recchia, T. Cathomen and M. A. Goncalves Differential integrity of TALE nuclease genes following adenoviral and lentiviral vector gene transfer into human cells Nucleic acids research 41 (2012) e63-e63.
  57. B. Minkenberg, M. Wheatley and Y. Yang CRISPR/Cas9-enabled multiplex genome editing and its application. in: Progress in molecular biology and translational science (Ed.^, Eds.), Elsevier, 2017, 111-132.
  58. K. Chen and C. Gao Targeted genome modification technologies and their applications in crop improvements Plant cell reports 33 (2014) 575-583.
  59. G. Zhang, Y. Lin, X. Qi, L. Li, Q. Wang and Y. Ma TALENs-assisted multiplex editing for accelerated genome evolution to improve yeast phenotypes ACS synthetic biology 4 (2015) 1101-1111.
  60. G. M. Masters Renewable and efficient electric power systems, John Wiley & Sons, 2013.
  61. W. Jiang, D. Bikard, D. Cox, F. Zhang and L. A. Marraffini RNA-guided editing of bacterial genomes using CRISPR-Cas systems Nature biotechnology 31 (2013) 233.
  62. G. Gasiunas, R. Barrangou, P. Horvath and V. Siksnys Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria Proceedings of the National Academy of Sciences 109 (2012) E2579-E2586.
  63. K. M. Esvelt, P. Mali, J. L. Braff, M. Moosburner, S. J. Yaung and G. M. Church Orthogonal Cas9 proteins for RNA-guided gene regulation and editing Nature methods 10 (2013) 1116.
  64. Z. Hou, Y. Zhang, N. E. Propson, S. E. Howden, L.-F. Chu, E. J. Sontheimer and J. A. Thomson Efficient genome engineering in human pluripotent stem cells using Cas9 from Neisseria meningitidis Proceedings of the National Academy of Sciences 110 (2013) 15644-15649.
  65. V. K. Shukla, Y. Doyon, J. C. Miller, R. C. DeKelver, E. A. Moehle, S. E. Worden, J. C. Mitchell, N. L. Arnold, S. Gopalan and X. Meng Precise genome modification in the crop species Zea mays using zinc-finger nucleases Nature 459 (2009) 437.
  66. G. Aad, B. Abbott, J. Abdallah, O. Abdinov, R. Aben, M. Abolins, O. AbouZeid, H. Abramowicz, H. Abreu and R. Abreu Combined Measurement of the Higgs Boson Mass in p p Collisions at s= 7 and 8 TeV with the ATLAS and CMS Experiments Physical review letters 114 (2015) 191803.
  67. F. Fauser, S. Schiml and H. Puchta Both CRISPR/Cas-based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana The Plant Journal 79 (2014) 348-359.
  68. Y. Zhang, Z. Liang, Y. Zong, Y. Wang, J. Liu, K. Chen, J.-L. Qiu and C. Gao Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA Nature communications 7 (2016) 12617.
  69. S. Kim, D. Kim, S. W. Cho, J. Kim and J.-S. Kim Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins Genome research 24 (2014) 1012-1019.
  70. T. Sprink, D. Eriksson, J. Schiemann and F. Hartung Regulatory hurdles for genome editing: process-vs. product-based approaches in different regulatory contexts Plant cell reports 35 (2016) 1493-1506.
  71. M. Malnoy, R. Viola, M.-H. Jung, O.-J. Koo, S. Kim, J.-S. Kim, R. Velasco and C. Nagamangala Kanchiswamy DNA-free genetically edited grapevine and apple protoplast using CRISPR/Cas9 ribonucleoproteins Frontiers in plant science 7 (2016) 1904.
  72. J. W. Woo, J. Kim, S. I. Kwon, C. Corvalán, S. W. Cho, H. Kim, S.-G. Kim, S.-T. Kim, S. Choe and J.-S. Kim DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins Nature biotechnology 33 (2015) 1162.
  73. S. Svitashev, C. Schwartz, B. Lenderts, J. K. Young and A. M. Cigan Genome editing in maize directed by CRISPR-Cas9 ribonucleoprotein complexes Nature communications 7 (2016) 13274.
  74. Z. Liang, K. Chen, T. Li, Y. Zhang, Y. Wang, Q. Zhao, J. Liu, H. Zhang, C. Liu and Y. Ran Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes Nature communications 8 (2017) 14261.
  75. A. Latorre, A. Latorre and íƒ. l. Somoza Modified RNAs in CRISPR/Cas9: An old trick works again Angewandte Chemie International Edition 55 (2016) 3548-3550.
  76. S. Ma, J. Chang, X. Wang, Y. Liu, J. Zhang, W. Lu, J. Gao, R. Shi, P. Zhao and Q. Xia CRISPR/Cas9 mediated multiplex genome editing and heritable mutagenesis of BmKu70 in Bombyx mori Scientific reports 4 (2014) 4489.
  77. X. Wang, Y. Wang, X. Wu, J. Wang, Y. Wang, Z. Qiu, T. Chang, H. Huang, R.-J. Lin and J.-K. Yee Unbiased detection of off-target cleavage by CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors Nature biotechnology 33 (2015) 175.
  78. S. S. Zaidi, M. M. Mahfouz and S. Mansoor CRISPR-Cpf1: A New Tool for Plant Genome Editing Trends Plant Sci 22 550-553.
  79. K. Xie, B. Minkenberg and Y. Yang Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system Proceedings of the National Academy of Sciences 112 (2015) 3570-3575.
  80. F. Fauser, S. Schiml and H. Puchta Both CRISPR/Cas"based nucleases and nickases can be used efficiently for genome engineering in Arabidopsis thaliana The Plant Journal 79 (2014) 348-359.
  81. L. G. Lowder, J. W. Paul, N. J. Baltes, D. F. Voytas, Y. Zhang, D. Zhang, X. Tang, X. Zheng, T.-F. Hsieh and Y. Qi A CRISPR/Cas9 toolbox for multiplexed plant genome editing and transcriptional regulation Plant physiology (2015) pp. 00636.02015.
  82. J. P. Guilinger, D. B. Thompson and D. R. Liu Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification Nature biotechnology 32 (2014) 577.
  83. J. You, L. Dou, K. Yoshimura, T. Kato, K. Ohya, T. Moriarty, K. Emery, C.-C. Chen, J. Gao and G. Li A polymer tandem solar cell with 10.6% power conversion efficiency Nature communications 4 (2013) 1446.
  84. H. Ledford US regulation misses some GM crops Nature 500 (2013) 389-390.
  85. N. Podevin, H. V. Davies, F. Hartung, F. Nogue and J. M. Casacuberta Site-directed nucleases: a paradigm shift in predictable, knowledge-based plant breeding Trends in biotechnology 31 (2013) 375-383.
  86. K. Pauwels, N. Podevin, D. Breyer, D. Carroll and P. Herman Engineering nucleases for gene targeting: safety and regulatory considerations New biotechnology 31 (2014) 18-27.
  87. H. Kim and J.-S. Kim A guide to genome engineering with programmable nucleases Nature Reviews Genetics 15 (2014) 321-334.
  88. J.-S. Lee, S.-J. Kwak, J. Kim, A.-K. Kim, H. M. Noh, J.-S. Kim and K. Yu RNA-guided genome editing in Drosophila with the purified Cas9 protein G3: Genes, Genomes, Genetics 4 (2014) 1291-1295.
  89. J. A. Zuris, D. B. Thompson, Y. Shu, J. P. Guilinger, J. L. Bessen, J. H. Hu, M. L. Maeder, J. K. Joung, Z.-Y. Chen and D. R. Liu Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo Nature biotechnology 33 (2015) 73.
  90. E. Waltz Tiptoeing around transgenics Nature Biotechnology 30 (2012) 215-217.
  91. H. D. Jones Regulatory uncertainty over genome editing Nature plants 1 (2015) 14011.
  92. S. Martin-Ortigosa, D. J. Peterson, J. S. Valenstein, V. S.-Y. Lin, B. G. Trewyn, L. A. Lyznik and K. Wang Mesoporous silica nanoparticle-mediated intracellular Cre protein delivery for maize genome editing via loxP site excision Plant physiology 164 (2014) 537-547.
  93. S. P. Jensen, V. J. Febres and G. A. Moore Cell penetrating peptides as an alternative transformation method in citrus Journal of Citrus Pathology 1 (2014) 276.
  94. Y. Wang, X. Cheng, Q. Shan, Y. Zhang, J. Liu, C. Gao and J.-L. Qiu Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew Nature biotechnology 32 (2014) 947.
  95. A. Piatek, Z. Ali, H. Baazim, L. Li, A. Abulfaraj, S. Alí¢€Shareef, M. Aouida and M. M. Mahfouz RNA-guided transcriptional regulation in planta via synthetic dCas9-based transcription factors Plant biotechnology journal 13 (2015) 578-589.
  96. L. G. Lowder, D. Zhang, N. J. Baltes, J. W. Paul, 3rd, X. Tang, X. Zheng, D. F. Voytas, T. F. Hsieh, Y. Zhang and Y. Qi A CRISPR/Cas9 Toolbox for Multiplexed Plant Genome Editing and Transcriptional Regulation Plant Physiol 169 (2015) 971-985.
  97. N. J. B. Tomáš Čermák, Radim Čegan, Yong Zhang and Daniel F. Voytas High-frequency, precise modification of the tomato genome Genome biology 16 (2015) 232.
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