IDT offers the components for genome editing through both their CRISPR-Cas9 system and CRISPR-Cpf1 system. The individual components can be ordered separately or as an entire system.

Benefits

  • Achieve higher efficiency genome editing than other methods by using Alt‑R® CRISPR RNAs, which are optimized, nuclease resistant, and individually quality controlled
  • Avoid toxicity or innate immune response activation by using Alt-R CRISPR ribonucleoproteins, instead of in vitro transcribed CRISPR nuclease mRNA and sgRNAs
  • Select from 2 CRISPR systems based on Cas9 or Cpf1 endonuclease for increased target site options
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A comparison of the two systems:

 Cas9 systemCpf1 system
ApplicationsGeneral genome editing• For species with AT-rich genomes
• For regions with limiting design space for use of the CRISPR-Cas9 system
Ribonucleoprotein components• crRNA
• tracrRNA
• Cas9 endonuclease
• crRNA
• Cpf1 endonuclease
crRNA• Native: 42 nt
• Alt-R: 35–36 nt (36 nt recommended)
• Native: 42–44 nt
• Alt-R: 40–44 nt (41 nt recommended)
tracrRNA• Native: 89 nt
• Alt-R: 67 nt
(not applicable)
CRISPR enzyme• Class 2, Cas type II
• M.W.*: 163,700 g/mol
• Endonuclease domains: RuvC-like and HNH
• Class 2, Cas type V
• M.W.*: 157,900 g/mol
• Endonuclease domain: RuvC-like only
Double-stranded DNA cleavage• Blunt ended cut 3 bases upstream of the protospacer sequence
• PAM site often destroyed during genome editing
• 5′ overhanging cut on the 5′ side of the protospacer sequence
• PAM site may be preserved after genome editing
PAM sequence†NGGTTTV
Current recommendations for Alt-R® RNP delivery• Lipid-mediated transfection
• Electroporation ± Alt-R® enhancer
• Microinjection
• Electroporation with Alt-R® enhancer
• Microinjection