CRISPR/Cas9 for Mutagenesis in Rice

Methods Mol Biol. 2019:1864:279-293. doi: 10.1007/978-1-4939-8778-8_19.

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.

Keywords: Agrobacterium-mediated rice transformation; CRISPR/Cas9; Genome editing; Rice; Targeted mutagenesis.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Agrobacterium tumefaciens / genetics
  • CRISPR-Cas Systems / genetics*
  • Gene Editing / instrumentation
  • Gene Editing / methods*
  • Genetic Vectors / genetics
  • Genome, Plant / genetics
  • Mutagenesis*
  • Oryza / genetics*
  • Plants, Genetically Modified / genetics*
  • RNA, Guide, CRISPR-Cas Systems / genetics
  • Tissue Culture Techniques / instrumentation
  • Tissue Culture Techniques / methods

Substances

  • RNA, Guide, CRISPR-Cas Systems