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
A comparison of the two systems:
| Cas9 system | Cpf1 system | |
|---|---|---|
| Applications | General 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† | NGG | TTTV |
| Current recommendations for Alt-R® RNP delivery | • Lipid-mediated transfection • Electroporation ± Alt-R® enhancer • Microinjection | • Electroporation with Alt-R® enhancer • Microinjection |