In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni (original) (raw)

“…Other natural CRISPR nucleases shown to function efficiently in mammalian cells include Staphylococcus aureus Cas9 (SaCas9) 10 , Acidaminococcus sp. Cpf1 11 , Lachnospiraceae bacterium Cpf1 11 , Campylobacter jejuni Cas9 12 , Streptococcus thermophilus Cas9 13 , and Neisseria meningitides Cas9 14 . None of these mammalian cell-compatible CRISPR nucleases, however, offer a PAM that occurs as frequently as that of SpCas9.…”

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“…Other natural CRISPR nucleases shown to function efficiently in mammalian cells include Staphylococcus aureus Cas9 (SaCas9) 10 , Acidaminococcus sp. Cpf1 11 , Lachnospiraceae bacterium Cpf1 11 , Campylobacter jejuni Cas9 12 , Streptococcus thermophilus Cas9 13 , and Neisseria meningitides Cas9 14 . None of these mammalian cell-compatible CRISPR nucleases, however, offer a PAM that occurs as frequently as that of SpCas9.…”

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“…The two Type II-C Cas9 orthologs (NmeCas9 and CjeCas9) that have been validated for 683 mammalian genome editing and assessed for genome-wide specificity (Lee et al 2016;Kim et al 2017) 684…”

Section: Truncated Sgrnas Reduce Off-target Cleavage By Nmecas9 571mentioning

“…Once a nuclease is chosen, it is important to ensure that gRNA target sequences are designed using the optimal length for that nuclease. For example, 20 bp is typically used for SpCas9 (Hsu et al, 2013), 22 bp for CjCas9 (Kim et al, 2017b), and 21–23 bp for SaCas9 (Ran et al, 2015). …”

Section: Library Designmentioning