ABM CRISPR

CRISPR/Cas9 allows for highly specific genomic modification and the silencing of genes of interest. This versatile system requires co-expression of two distinct components: (1) a nuclease, Cas9, and (2) a target-specific  single guide RNA (sgRNA). Streptococcus pyogenes Cas9 interrogates the genome for sequences complementary to the 20 nucleotide target region of the sgRNA and adjacent to the protospacer-adjacent motif (PAM) “5’-NGG”. The Cas9 nuclease introduces a double strand break, which is then repaired by a highly error-prone process called Non-Homologous End Joining (NHEJ). This can result in a frameshift insertion or deletion (InDel), thus effectlively silencing the gene.