CRISPR directed evolution of the spliceosome for resistance to splicing inhibitors

Genome Biol. 2019 Apr 30;20(1):73. doi: 10.1186/s13059-019-1680-9.


Increasing genetic diversity via directed evolution holds great promise to accelerate trait development and crop improvement. We developed a CRISPR/Cas-based directed evolution platform in plants to evolve the rice (Oryza sativa) SF3B1 spliceosomal protein for resistance to splicing inhibitors. SF3B1 mutant variants, termed SF3B1-GEX1A-Resistant (SGR), confer variable levels of resistance to splicing inhibitors. Studies of the structural basis of the splicing inhibitor binding to SGRs corroborate the resistance phenotype. This directed evolution platform can be used to interrogate and evolve the molecular functions of key biomolecules and to engineer crop traits for improved performance and adaptation under climate change conditions.

Keywords: CRISPR/Cas9; Directed evolution; Genome engineering; Herbicide resistance; Herboxidiene; Pladienolide B; SF3B complex; SF3B1; Spliceosome; Spliceostatin A; Splicing modulators.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Clustered Regularly Interspaced Short Palindromic Repeats*
  • Evolution, Molecular*
  • Fatty Alcohols
  • Genetic Techniques*
  • Oryza / genetics*
  • Plant Proteins / genetics
  • Protein Domains
  • Pyrans
  • Spliceosomes*


  • Fatty Alcohols
  • Plant Proteins
  • Pyrans
  • herboxidiene