Generation of Targeted Point Mutations in Rice by a Modified CRISPR/Cas9 System

CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated Cas9 endonuclease)-mediated genome editing has revolutionized biological research and crop improvement because of its specificity, simplicity, and versatility (reviewed in Komor et al., 2016a). Editing a gene by CRISPR/Cas9 only has three requirements: (1) expression of the nuclear localized Cas9 protein; (2) production of a guide RNA (gRNA) molecule, whose first 20 nucleotides are complementary to the target gene; (3) the NGG PAM site that is located immediately adjacent to the 3′ end of the target sequence (Figure 1A).