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CRISPR/Cas9 for Mutagenesis in Rice

  • Si Nian Char
  • Riqing Li
  • Bing Yang
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1864)

Abstract

CRISPR/Cas9 (clustered regularly interspaced short palindromic repeat/CRISPR-associated protein 9) provides a workhorse for genome editing biotechnology. CRISPR/Cas9 tailored for enabling genome editing has been extensively interrogated and widely utilized for precise genomic alterations in eukaryotic organisms including in plant species. The technology holds the great promise to better understand gene functions, elucidate networks, and improve the performance of crop plants such as increasing grain yields, improving nutritional content, and better combating the biotic and abiotic stresses. Various methods or protocols specific for different plant species have been established. Here, we present a CRISPR/Cas9-mediated genome editing protocol in rice, including detailed information about single-guide RNA design, vector construction, plant transformation, and mutant screening processes.

Key words

CRISPR/Cas9 Genome editing Rice Agrobacterium-mediated rice transformation Targeted mutagenesis 

Notes

Acknowledgments

The authors acknowledge the funding support from National Science Foundation (IOS-1238189 to BY), the National Institute of Food and Agriculture of the US Department of Agriculture (2014-67013-21720 to BY).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Genetics, Development and Cell BiologyIowa State UniversityAmesUSA

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