rNA-guided editing of bacterial genomes using crisPr-cas systems

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CRISPR/Cas9 systems have off-target activity with insertions or deletions between target DNA and guide RNA sequences (2014, unknown): “Mismatches in the PAM sequence are less tolerated, although Cas9 also recognizes an alternative NAG PAM with low frequency (20,23,25).”
CRISPR-P: a web tool for synthetic single-guide RNA design of CRISPR-system in plants. (2014, methodology): “Until now, the CRISPR/Cas system has been successfully applied to efficient genome editing in bacteria, animals and plants (Jiang et al., 2013; Mali et al., 2013; Xie and Yang, 2013).”
Multigene Editing in the Escherichia coli Genome via the CRISPR-Cas9 System (2015, methodology): “To reduce the off-target effects of Cas9 in this study, an N20 sequence was selected to ensure the last 12 bp was highly specific for the targets (24).”
CRISPR/Cas9 in Genome Editing and Beyond. (2016, background): “Its natural endonuclease activity has been co-opted for sequence-specific editing of the genome in a wide range of organisms, including bacteria (31), fungi (32), plants (33, 34), and animals (5–7, 9, 10, 35, 36).”
Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells (2014, unknown): “Base pairing in the first 10-12 nucleotides adjacent to PAM (defined as the “seed”) was found to be generally more important than pairing in the rest of the guide region 6,8,16,33 .”