Advances in CRISPR/Cas technologies and their application in plants

Abstract

Discovery of the Clustered Regularly Interspaced Short Palindromic Repeats / CRISPR-associated (CRISPR/Cas) system and its repurposing into a powerful genome editing tool has revolutionized genome engineering and generated excitement for innovative breeding technology. CRISPR/Cas can perform genetic operations such as targeted insertion, deletion and replacement of genes in plants by its simple two-component system comprising a Cas protein and a guide RNA. Here, we focus on the recent advances in CRISPR/Cas technologies that are available for genome editing, among which Cas9, Cas12, and Cas13 systems are widely used in the areas of botany and agriculture. We also describe the new high-precision genome editing tools, base editors and prime editors that are derived from the Cas9 system and beyond for altering the genome in living cells, without generating double-stranded breaks in DNA or requiring a donor. In addition, we summarize the differences between the different CRISPR/Cas systems and their broad applications in plants. We also discuss the challenges facing the use of CRISPR/Cas technologies and the future directions of CRISPR/Cas systems in plant genome editing.