The Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system is the latest RNA-guided endonuclease tool in genome editing which allows for very specific genomic disruption and replacement. ;;The Cas9 nuclease serves to unwind the genomic DNA duplex next to conserved protospacer adjacent motifs (PAMs) and homes in on its target sequence, which is recognized by a complementary single-guide RNA. The resulting double-stranded break gets repaired by the non-homologous end joining (NHEJ) pathway, leading to a disruption in the open reading frame of the targeted gene. Alternatively, by supplying a suitable repair template, virtually any desired point mutation can be introduced at the break point via homology-directed repair (HDR).;;The Cas9 nuclease from the bacteria Staphylococcus aureus, abbreviated saCas9, is gaining popularity as an alternative to spCas9 due to its relatively smaller size. The saCas9 PAM sequence is 5’-NNGRRN (preferably 5’-NNGRRT).
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