Reengineering CRISPR–cas for genome editing

+ 22 chNA trachNA Sequence of guide RNA gene designed to target locus of interest poNlA llI NIA Ill promoter prgfnoter Guide RNA C359 _ . Transcription Transcnptlon and translation Ca59 I gRNA is a fusion of the chNA + trachNA I I Cleavage dsDNA target in genome :- I I Donor DNA with desired sequence NHEJ repair of double-strand l l breaks results in deletions of l I I one or more nucleotides, I I or sometimes insertion of DNA ‘ Mutation, 99, Edited genomic derived from elsewhere in l deletion locus the genome l I Non—homologous Homologous end ioining recombination