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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.

Alt-R™ CRISPR-Cas9 System

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

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Alt-R™ CRISPR-Cas12a (Cpf1) System

For additional target sites or for targeting AT-rich regions, use the CRISPR-Cas12a system in electroporation experiments. Protospacer adjacent motif (PAM) = TTTV. The Alt-R Cas12a (Cpf1) Ultra also can recognize many TTTT PAM sites in addition to TTTV motifs, increasing target range for genome editing studies.

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Alt-R™ CRISPR nucleases

Recombinant, high-purity Cas9 and Cas12a (Cpf1) endonucleases for genome editing experiments.

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Alt-R CRISPR Custom Guide RNAs

Custom guide RNAs ideal for prime editing (pegRNA) projects, CRISPR-Cas13 applications, and most alternative CRISPR-Cas systems.

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Alt-R CRISPR Custom gRNA Libraries

Custom arrayed synthetic guide RNA libraries are a great way to accomplish CRISPR knockout screens. They accelerate your research by avoiding the cloning and sequencing associated with lentiviral screens while also integrating into existing automation workflows. 

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Alt-R™ HDR Donor Oligos

Single-stranded DNA oligos specifically built for your homology‑directed repair (HDR) experiments. Designed and optimized through extensive laboratory testing, they are ideal for introducing point mutations and short insertions.

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Alt-R HDR Donor Blocks

Double-stranded DNA fragments specifically built for your homology-directed repair (HDR) experiments. IDT dsDNA templates offer a cost-effective, high-fidelity option to reduce the amount of downstream screening via high HDR rates and low unintended blunt integrations. Alt-R HDR Donor Blocks are ideal for making large insertions or genomic changes.

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rhAmpSeq™ CRISPR Analysis System

An end-to-end solution to design, deploy, and analyze next generation sequencing data for on- and off-target interrogation after your CRISPR experiment.

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Alt-R™ Genome Editing Detection Kit

T7 endonuclease I (T7EI) mismatch cleavage assay for detection of on-target editing, known off-target events, and estimation of genome editing efficiency in cultured cells.

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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 sequence†NGG
  • 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.

*RUO—For research use only. Not for use in diagnostic procedures. Unless otherwise agreed to in writing, IDT does not intend for these products to be used in clinical applications and does not warrant their fitness or suitability for any clinical diagnostic use. Purchaser is solely responsible for all decisions regarding the use of these products and any associated regulatory or legal obligations.

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