Fig. 1

CRISPR-enabled plant directed evolution systems. Clustered regularly interspaced palindromic repeat (CRISPR) libraries can be constructed through ribonucleoprotein (RNP) assembly or cloning guide RNA (gRNA) library into plasmids. CRISPR libraries can be delivered into plants through Agrobacterium-mediated transformation, viral vectors, particle bombardment, or polyethylene glycol (PEG)-mediated transformation. Plant cells, explants used for regeneration, or whole plants can be used as evolution platforms. Selected mutants can be sequenced to reveal the genetic changes. Protein and phenotypic characterization can be carried out to study the evolution consequences