Method for modifying target nucleic acid in genome of cell

A method for modifying a target nucleic acid in the genome of a cell, the method comprising:

• incubating a cell comprising a target nucleic acid;
• a polynucleotide encoding a clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nuclease or a variant thereof;
• and a guide RNA,
• wherein the target nucleic acid comprises a protospacer adjacent motif (PAM) and a target sequence complementary to the guide RNA,
• the PAM consists of a sequence selected from the group consisting of 5′-NA-3′, 5′-NTG-3′, 5′-GTH-3′, 5′-VCGD-3′, 5′-TCGDG-3′, 5′-GGT-3′, 5′-BGC-3′, 5′-RTG-3′, 5′-NYGG-3′, 5′-AGCCA-3′, 5′-CTGCA-3′, 5′-TGTCA-3′, 5′-GCGCA-3′, 5′-GCGCT-3′, 5′-GCGTT-3′, 5′-AGCCC-3′, 5′-CGTCA-3′, 5′-GCGTA-3′, 5′-AGCCG-3′, 5′-GCGTC-3′, 5′-AGTCA-3′, 5′-AGCAC-3′, and 5′-AGCTC-3′,
• the target nucleic acid can be recognized by a complex of the Cas nuclease or a variant thereof with the guide RNA, and
• the complex of the Cas nuclease or a variant thereof with the guide RNA modifies the target nucleic acid sequence-specifically.