CRISPR/Sc++ -mediated genome editing in rice

J Integr Plant Biol. 2021 Sep;63(9):1606-1610. doi: 10.1111/jipb.13166.

Abstract

Streptococcus canis Cas9 (ScCas9) is an RNA-guided endonuclease with NNG protospacer adjacent motif (PAM) specificity whose genome-editing activity in rice is locus-dependent. Here we investigated the performance of a ScCas9 variant named Sc++ at different NNG PAM sites in the rice genome; Sc++ harbors a T1227K mutation and the substitution of a positively charged loop (residues 367-376). Sc++ nuclease achieved broader genome editing compared to the original ScCas9, and its nickase improved targeted base editing in transgenic rice plants. Using the high-efficiency adenine base editor rBE73b, we generated many new OsGS1 alleles suitable for screening of rice germplasm for potential herbicide resistance in the future. The CRISPR/Sc++ system expands the genome-editing toolkit for rice.

Keywords: Sc++; base editing; genome editing; rice.

Publication types

  • Evaluation Study

MeSH terms

  • CRISPR-Associated Protein 9 / genetics*
  • CRISPR-Cas Systems*
  • Gene Editing / methods*
  • Genome, Plant
  • Oryza / genetics*
  • Streptococcus / genetics

Substances

  • CRISPR-Associated Protein 9

Supplementary concepts

  • Streptococcus canis