CRISPR/Cas9-Mediated Editing of Genes Encoding rgs-CaM-like Proteins in Transgenic Potato Plants

Methods Mol Biol. 2019:2028:153-165. doi: 10.1007/978-1-4939-9635-3_9.

Abstract

A tobacco calmodulin-like protein, rgs-CaM, has been shown to interact with viruses in a variety of ways; it contributes to geminivirus infections but is also involved in primed immunity to the cucumber mosaic virus. Sequence similarity searches revealed several calmodulin-like proteins similar to rgs-CaM (rCML) in Arabidopsis and other Solanaceae plants, including potato (Solanum tuberosum). To analyze the functions of each rCML, mutations were introduced into potato rCMLs using the CRISPR/Cas9 system. Here, we describe our protocol of the CRISPR/Cas9-mediated targeted mutagenesis in stably transformed potato plants.

Keywords: CRISPR/Cas9; Calmodulin-like protein; Potato plants; Targeted mutagenesis; rgs-CaM.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • CRISPR-Cas Systems*
  • DNA Mutational Analysis
  • Gene Editing*
  • Gene Targeting
  • Genetic Vectors / genetics
  • Mutagenesis
  • Phenotype
  • Plant Proteins / genetics*
  • Plants, Genetically Modified*
  • Solanum tuberosum / genetics*
  • Transformation, Genetic

Substances

  • Plant Proteins
  • rgs-CaM protein, Nicotiana tabacum