Icons Library_Wave_180x180_CRISPR

CRISPR genome editing

The Alt-R CRISPR Systems were developed through comprehensive research on each component of the CRISPR-driven technology that generates a double-stranded break critical for gene disruption and DNA insertion by homologous recombination.

A complete workflow solution, from design to analysis.

Overview

Use of the CRISPR (clustered regularly interspaced short palindromic repeats) and associated Cas9 enzyme for genome editing has been a major technological breakthrough, making genome modification in cells or organisms faster, more efficient, and more robust than previous genome editing methods. The Alt-R CRISPR-Cas9 System is an optimized genome editing solution for producing on-target, double-stranded DNA breaks.

We have also developed an alternative Alt-R CRISPR-Cas12a (Cpf1) System to open up CRISPR editing to additional areas in genomes.

Quick comparison of CRISPR genome editing using Cas9 vs. Cas12a (Cpf1)

Figures showing difference between Cas9 and Cas12 enzymes
 Cas9 systemCas12a 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
  • gRNA options:
    1. crRNA and tracrRNA
    2. sgRNA
  • Cas9 endonuclease
  • crRNA
  • Cas12a endonuclease
Variants
  • Wild-type
  • HiFi
  • Nickases (H840A and D10A)
  • Wild-type
  • Ultra (Improved performance)
Cas9 crRNA:tracrRNA (option 1)

crRNA

  • Native: 42 nt
  • Alt-R: 35–36 nt (36 nt recommended)

tracrRNA

  • Native: 89 nt
  • Alt-R: 67 nt
Cas9 sgRNA (option 2)
  • Alt-R: 99–100 nt (100 nt recommended)
Cas12a crRNA
  • Native: 42–44 nt
  • Alt-R: 40–44 nt (41 nt recommended)
CRISPR enzyme
  • Class 2, Cas type II
  • M.W.*: 162,200 g/mol
  • Endonuclease domains: RuvC-like and HNH
  • Class 2, Cas type V
  • M.W.*: 156,400 g/mol
  • Endonuclease domain: RuvC-like only
DNA cleavage
  • Wild-type and HiFi: Blunt-ended cut 3 bases upstream of the protospacer sequence
  • D10A nickase with paired gRNAs: 5′ overhang
  • H840A nickase with paired gRNAs: 3′ overhang
  • 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 sequenceNGG
  • TTTV for Cas12a V3
  • TTTN for Cas12a Ultra
Current recommendations for Alt-R RNP delivery
  • Lipid-mediated transfection
  • Electroporation (Alt-R enhancer recommended)
  • Microinjection
  • Electroporation (Alt-R enhancer recommended)
  • Microinjection

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

IDT does not sell gene therapy kits and nothing sold by IDT should be construed as a gene therapy kit. Customers should not use any IDT products for self-administration.

RUO22-1177_001