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Biolistic Delivery of Programmable Nuclease (CRISPR/Cas9) in Bread Wheat

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Biolistic DNA Delivery in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2124))

Abstract

The discovery of site-specific programmable nucleases has led to a major breakthrough in the area of genome editing. In the past few years, CRISPR/Cas system has been utilized for genome editing of a large number of crops including cereals like wheat, rice, maize, and barley. In terms of consumption, wheat is second only to rice as the most important crop of the world. In the present chapter, we describe biolistic delivery method of ribonucleoprotein (RNP) complexes of programmable nuclease (CRISPR/Cas9) for targeted genome editing and selection-free screening of transformants in wheat. The method not only overcomes the problem of random integration into the genome but also reduces the off-targets. Besides the step-by-step protocol, plausible challenges and ways to overcome them are also discussed. By using the described method of biolistic delivery of CRISPR/Cas9 in plant systems, genome-edited plants can be identified within 11 weeks.

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Author information

Authors and Affiliations

  1. National Agri-Food Biotechnology Institute (NABI), Mohali, Punjab, India

    Abhishek Bhandawat, Vinita Sharma, Vikas Rishi & Joy K. Roy

Authors
  1. Abhishek Bhandawat
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  2. Vinita Sharma
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  3. Vikas Rishi
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  4. Joy K. Roy
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Corresponding author

Correspondence to Joy K. Roy .

Editor information

Editors and Affiliations

  1. Department of Plant and Environmental Sciences, School of Health Research, Pee Dee Research and Education Center, Clemson University, Florence, SC, USA

    Sachin Rustgi

  2. Department of Genetics and Biochemistry, Clemson University, Clemson, SC, USA

    Hong Luo

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Bhandawat, A., Sharma, V., Rishi, V., K. Roy, J. (2020). Biolistic Delivery of Programmable Nuclease (CRISPR/Cas9) in Bread Wheat. In: Rustgi, S., Luo, H. (eds) Biolistic DNA Delivery in Plants. Methods in Molecular Biology, vol 2124. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0356-7_17

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  • DOI: https://doi.org/10.1007/978-1-0716-0356-7_17

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0355-0

  • Online ISBN: 978-1-0716-0356-7

  • eBook Packages: Springer Protocols

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