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Production of Virus-Resistant Plants Through CRISPR-Cas Technology

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Characterization of Plant Viruses

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Abstract

Clustered regularly interspaced short palindromic repeats (CRISPR) are widely found in bacterial and archaeal genomes as a defence mechanism against invading viruses and plasmids. The CRISPR locus consists of segments of prokaryotic DNA with short repetitions of base sequences. Each repetition is followed by short segments of ‘spacer DNA’ from earlier exposed bacterial virus or plasmid. CRISPR spacers recognize and cut these exogenous genetic elements in a manner analogous to RNA interference in eukaryotic organisms. The CRISPR interference technique has enormous potential application, including altering the germline of humans, animals, other organisms and plants. The organism’s genome can be cut at any desired location by delivering the Cas9 protein and guide RNAs into a cell. Thus CRISPR-Cas system represents a powerful tool in developing resistance to DNA and RNA plant viruses by editing and inserting novel traits precisely at chosen loci into plants and offers enormous potential in classical breeding. It has opened new way to get virus-resistant plants either by directly targeting and cutting the viral genome, or by modifying the host genome itself to introduce viral immunity. In this chapter, recent progress demonstrating the efficacy of the CRISPR/Cas technology against DNA and RNA plant viruses is discussed.

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References

  • Ali Z, Abulfaraj A, Idris A, Ali S, Tashkandi M, Mahfouz MM (2015) CRISPR/Cas9-mediated viral interference in plants. Genome Biol 16:238

    Article  Google Scholar 

  • Ali Z, Ali S, Tashkandi M, Zaidi SS, Mahfouz MM (2016) CRISPR/Cas9-mediated immunity to geminiviruses: differential interference and evasion. Sci Rep 6(26):9–12

    Google Scholar 

  • Aman R, Ali Z, Butt H, Mahas A, Aljedaani F, Zuhaib Khan M, Ding S, Mahfouz M (2018) RNA virus interference via CRISPR/Cas13a system in plants. Genome Biol 19:1

    Article  Google Scholar 

  • Baltes NJ, Hummel AW, Konecna E, Cegan R, Bruns AN, Bisaro DM (2015) Conferring resistance to geminiviruses with the CRISPR–Cas prokaryotic immune system. Nat Plants 1:15145

    Article  CAS  Google Scholar 

  • Chandrasekaran J, Brumin M, Wolf D, Leibman D, Klap C, Pearlsman M (2016) Development of broad virus resistance in non-transgenic cucumber using CRISPR/Cas9 technology. Mol Plant Pathol 17:1140–1153

    Article  CAS  Google Scholar 

  • Doudna JA, Charpentie E (2014) The new frontier of genome engineering with CRISPR-Cas9. Science 346(6213):1258096

    Article  Google Scholar 

  • Fondong VN (2013) Geminivirus protein structure and function. Mol Plant Pathol 14:635–649

    Article  CAS  Google Scholar 

  • Hanley-Bowdoin L, Bejarano ER, Robertson D, Mansoor S (2013) Geminiviruses: masters at redirecting and reprogramming plant processes. Nat Rev Microbiol 11:777–788

    Article  CAS  Google Scholar 

  • Ishino Y, Shinagawa H, Makino K, Amemura M, Nakata A (1987) Nucleotide sequence of the IAP gene, responsible for alkaline phosphatase isozyme conversion in Escherichia coli, and identification of the gene product. J Bacteriol 169:5429–5433

    Article  CAS  Google Scholar 

  • Ji X, Zhang H, Zhang Y, Wang Y, Gao C (2015) Establishing a CRISPR-Cas-like immune system conferring DNA virus resistance in plants. Nat Plants 1:15144

    Article  CAS  Google Scholar 

  • Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E (2012) A programmable dual-RNA–guided DNA endonuclease in adaptive bacterial immunity. Science 337:816–821

    Article  CAS  Google Scholar 

  • Kanchiswamy CN (2016) DNA-free genome editing methods for targeted crop improvement. Plant Cell Rep 35:1469–1474

    Article  CAS  Google Scholar 

  • Khan MZ, Amin A, Hameed A, Mansoor S (2018) CRISPR–Cas13a: prospects for plant virus resistance. Cell 36(12):1207–1210

    CAS  Google Scholar 

  • Larson MH, Gilbert LA, Wang X, Lim WA, Weissman JS, Qi LS (2013) CRISPR interference (CRISPRi) for sequence-specific control of gene expression. Nat Protoc 8(11):2180–2196

    Article  CAS  Google Scholar 

  • Liu X, Wu S, Xu J, Sui C, Wei J (2017) Application of CRISPR/Cas9 in plant biology. Acta Pharm Sin 7:292–302

    Article  Google Scholar 

  • Macovei A, Sevilla NR, Cantos C, Jonson GB, Slamet-Loedin I, Čermák T, Voytas DF, Choi IR, Chadha-Mohanty P (2018) Novel alleles of rice eIF4G generated by CRISPR/Cas9-targeted mutagenesis confer resistance to Rice tungro spherical virus. Plant Biotechnol J 16(11):1918–1927

    Article  CAS  Google Scholar 

  • Pyott DE, Sheehan E, Molnar A (2016) Engineering of CRISPR/Cas9-mediated potyvirus resistance in transgene-free Arabidopsis plants. Mol Plant Pathol 17:1276–1288

    Article  CAS  Google Scholar 

  • Sanfacon H (2015) Plant translation factors and virus resistance. Viruses 7:3392–3419

    Article  CAS  Google Scholar 

  • Voytas DF, Gao C (2014) Precision genome engineering and agriculture: opportunities and regulatory challenges. PLoS Biol 12:e1001877

    Article  Google Scholar 

  • Woo JW, Kim J, Kwon SI, Corvalan C, Cho SW, Kim H et al (2015) DNA-free genome editing in plants with preassembled CRISPR-Cas9 ribonucleoproteins. Nat Biotechnol 33:1162–1164

    Article  CAS  Google Scholar 

  • Zhang T, Zheng Q, Yi X, An H, Zhao Y, Ma S et al (2018) Establishing RNA virus resistance in plants by harnessing CRISPR immune system. Plant Biotechnol J 16(8):1415–1423

    Article  CAS  Google Scholar 

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Bhat, A.I., Rao, G.P. (2020). Production of Virus-Resistant Plants Through CRISPR-Cas Technology. In: Characterization of Plant Viruses . Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0334-5_50

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  • DOI: https://doi.org/10.1007/978-1-0716-0334-5_50

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0333-8

  • Online ISBN: 978-1-0716-0334-5

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