CRISPR genome editing

For manipulating eukaryotic genomes

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
Related product

Alt-R™ Genome Editing Detection Kit—to monitor on-target CRISPR editing or estimate editing efficiency.


CRISPR nucleases

Convenient ordering of Cas9 or Cpf1 protein.

Alt-R™ CRISPR-Cas9 System

Get efficient CRISPR reagents based on the commonly used Streptococcus pyogenes Cas9 system for lipofection or electroporation experiments. Protospacer adjacent motif (PAM) = NGG.

Alt-R™ CRISPR-Cpf1 System

If you need additional target sites or are targeting AT-rich regions, use the Acidaminococcus sp. BV3LC CRISPR-Cpf1 system in electroporation experiments. Protospacer adjacent motif (PAM) = TTTN.


Quick comparison of CRISPR genome editing using Cas9 vs. Cpf1


cas9_cpf1_compare

 

  Cas9 system Cpf1 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 NGG TTTV
Current recommendations for Alt-R™ RNP delivery • Lipid-mediated transfection
• Electroporation ± Alt-R™ enhancer 
• Microinjection
• Electroporation with Alt-R™ enhancer
• Microinjection

* Molecular weight of Alt-R™ nuclease
N = any base; V = A, C, or G