Alt-R S.p Cas9 Nuclease 3NLS
Alt-R S.p. Cas9 Nuclease 3NLS enzyme is a high purity, recombinant S. pyogenes Cas9. The enzyme includes 1 N-terminal nuclear localization sequence (NLS) and 2 C-terminal NLSs, as well as a C-terminal 6-His tag. The S. pyogenes Cas9 enzyme must be combined with a crRNA and tracrRNA in order to produce a functional, target-specific editing complex. For the best editing, combine the Alt-R S.p. Cas9 Nuclease 3NLS enzyme with the optimized Alt-R CRISPR crRNA and tracrRNA in equimolar amounts.
Alt-R S.p. HiFi Cas9 Nuclease 3NLS
Alt-R S.p. HiFi Cas9 Nuclease offers improved specificity over wild-type Cas9, greatly reducing the risk of off-target cutting events. This Cas9 variant also preserves the high level of editing efficiency expected from a Cas9 nuclease, maintaining 90–100% on-target editing activity at most sites. For applications that are sensitive to off-target events, combining the Alt-R S.p. HiFi Cas9 Nuclease with the optimized Alt-R CRISPR-Cas9 crRNA and Alt-R CRISPR-Cas9 tracrRNA is highly recommended.
Alt-R S.p. Cas9 Nickases
Cas9 nickases allow specific cutting of only one strand at the DNA target site. Cuts to both strands of DNA are accomplished by using both the Alt-R S.p. Cas9 D10A and the Alt-R S.p. Cas9 H840A Nickase 3NLS, and targeting two neighboring Cas9 sites, functionally increasing the length of the recognition sequence from 20 to 40 bases. For more information about using Cas9 nickases, see the application note.
Alt-R A.s. Cpf1 Nuclease 2NLS
Alt-R A.s. Cpf1 Nuclease 2NLS enzyme is a high purity, recombinant Acidaminococcus sp. BV3L6 Cpf1. The enzyme includes 1 N-terminal nuclear localization sequence (NLS) and 1 C-terminal NLS, as well as 1 N-terminal V5 tag and 1 C-terminal 6-His tag. The Cpf1 enzyme must be combined with a crRNA to produce a functional, target-specific editing complex. For the best editing, combine Alt-R A.s. Cpf1 Nuclease 2NLS enzyme with optimized Alt-R CRISPR-Cpf1 crRNA in equimolar amounts.
Attention: Unlike S. pyogenes Cas9, which cleaves most NGG PAM sites to some degree, some of the tested TTTV sites show no cleavage by A.s. Cpf1 nuclease. We recommend using positive control crRNAs to establish that your cells can be edited by Cpf1. In addition, we suggest testing 3 or more crRNAs per target gene.