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. 2021 Aug 9;1(8):e207. doi: 10.1002/cpz1.207

Figure 3.

Figure 3

Locations of CRISPR/Cas9 cut sites relative to the position of the human NBS1 gene. (A) The NBS1 gene is located at Chr8: positions 91,015,482‐91,070,776 [Mar 2006 (NSB136/hg18)]. The 3′ and 5′ ends of the NBS1 (targeted genomic sequences) homologous to the TAR cloning vector hooks are marked in red (hook 1) and in green (hook 2), respectively. The cut sites of Cas9‐sgRNA‐A and Cas9‐sgRNA‐B complexes upstream of hook 1, and of Cas9‐sgRNA‐C and Cas9‐sgRNA‐D complexes downstream of hook 2, with their positions toward the hooks, are indicated. (B) Schematic representation of Cas9‐sgRNA binding to DNA. The CRISPR guide sequence is cloned into the 5′ end of the sgRNA molecule. The guide sequence is any 20‐bp DNA sequence upstream of an NGG, the Protospacer Adjacent Motif (PAM). The Cas9 cut site is 3 bp upstream of PAM. (C) Guide sequences used for the human NBS1 gene cloning are illustrated. This figure was adapted from Lee et al. (2015).