CRISPR-Cas-Mediated Gene Knockout in Tomato

Methods Mol Biol. 2020:2083:321-341. doi: 10.1007/978-1-4939-9952-1_25.


Loss-of-function mutants are crucial for plant functional genomics studies. With the advent of CRISPR-Cas genome editing, generating null alleles for one or multiple specific gene(s) has become feasible for many plant species including tomato (Solanum lycopersicum). An easily programmable RNA-guided Cas endonuclease efficiently creates DNA double-strand breaks (DSBs) at targeted genomic sites that can be repaired by nonhomologous end joining (NHEJ) typically leading to small insertions or deletions that can produce null mutations. Here, we describe how to utilize CRISPR-Cas genome editing to obtain stable tomato gene knockout lines.

Keywords: CRISRP-Cas; Gene knockout; Genome editing; Loss-of-function mutation; Null mutation; Site-directed mutagenesis; Solanaceae; Solanum lycopersicum; Targeted mutagenesis; Tomato.

MeSH terms

  • CRISPR-Cas Systems*
  • DNA End-Joining Repair
  • Gene Editing
  • Gene Knockout Techniques*
  • Gene Targeting
  • Genetic Vectors / genetics
  • Plants, Genetically Modified
  • RNA, Guide, CRISPR-Cas Systems
  • Solanum lycopersicum / genetics*
  • Workflow


  • RNA, Guide, CRISPR-Cas Systems