AccuTool™ gRNA & Cas9 protein (RNP)

Active RNA-Guided Endonuclease (aRGEN)) is a new generation of engineered nuclease based on the CRISPR-Cas9 system. It is an innovative tool for genome editing composed of Cas9 nuclease and gRNA.
By recognizing the target sequence (19~20 bp) and PAM sequence (5'-NGG-3'), aRGEN can easily edit any genomic regions. The target DNA region can be effectively cleaved during either in vivo or in vitro genome editing applications (knock-in/out, and locus deletion).

RNP Complex of gRNA and Cas9 Protein

The target-specific gRNA and Cas9 protein can form a stable ribonucleoprotein (RNP) complex, and further into a target-specific nuclease that can be active in both in vitro and in vivo. The complex can be used on your cell of interest that are difficult to be transfected like primary cells. Furthermore, after the transfection, it has a short lifetime in the cell, minimizing off-target effects. With this, AccuTool ™ gRNA & Cas9 protein (RNP) will provide the most effective and detailed methodologies.

• aRGEN-Lyophilized sgRNA Service
  AccuTool™ aRGEN service provides a custom-designed sgRNA for optimal efficiency and specificity.
  Furthermore, aRGEN positive controls targeting EGFP, CCR5 and HPRT1 genes are provided. They are ready-to-use products sent in a lyophilized form.
• Recombinant Cas9 protein Selection Guide (Figure 1)
  All the recombinant Cas9 proteins are shipped in solution form.
  

  Cas9 WT Protein	This protein is the most used Cas9 protein derived from Streptococcus pyogenes bacteria. It is an RNA-guided nuclease that creates a site-specific DSBs.
  Nickase (D10A) Cas9 Protein	This protein is a variant of Cas9 protein differing by a point mutation (D10A). It can create a single-stranded nick in each DNA strand, resulting in DSBs on the target DNA region. Each of the cleaved ends forms overhangs instead of blunt ends, allowing control of the integration and insertion process of the genes.
  Dead (D10A/H840A) Cas9 Protein	This protein is a variant of Cas9 protein differing by point mutations (D10A and H840A). It does not have endonuclease activity but still retains its ability to bind onto the target sequences. With this, dead (D10A/H840A) cas9 proteins can be used to control gene expression by forming a complex with transcriptional activators or repressors.
  Sniper Cas9 Protein	This protein is a variant of SpCas9 which lowered the off-target effect and enhanced the on-target effect (high-specificity, low off-target).
  Cyanine3-Cas9 Protein	The Cas9 WT and the dead (D10A/H840A) cas9 proteins are labeled with a fluorescent dye, Cyanine3, for customers to directly track their delivery to the target cells.

  

  

  Figure 1. Recombinant Cas9 protein

• Cas9 nuclease

  All the proteins and mRNA can be delivered in requested forms (Figure 2).

  gRNA-Cas9 protein RNP complex	- gRNA-Cas9 protein RNP complex can be delivered to the cell of interest that are difficult to be transfected with electroporation.  - Transcription or translation is not needed for Cas9 and gRNA expression. Use the complex directly for easier genome editing. - This product is recommended for researchers transfecting directly to a nucleus, using methods such as nucleofection and microinjection.	- These products do not affect cell-type-specific promoter activities. - These products pose no risk of random integration to the host genome.
  gRNA and Cas9 mRNA	- gRNA and Cas9 mRNA can be delivered to your cell of interest either by chemical transfection or electroporation. - Transcription is not needed. Use the product directly for easier genome editing.

   

  

  Figure 2. Cas9 nuclease

Features and Benefits

• High knock-out efficiency
  

  Low toxicity and high efficiency compared to its plasmid form. Reduced off-target effects with short lifetime in cells.

• Easy and simple to use
  

  Apply to the target cells without any sample preparation step. Create KO animals with embryo injection.

• Economical & Fast delivery
  

  Get your products quickly at a reasonable price.