AccuTool™ Kit

• AccuTool ™ CRISPR-Cas9 Starter Kit

Overview

AccuTool™ CRISPR-Cas9 Starter Kit is an all-in-one kit recommended for researchers starting plasmid CRISPR-Cas9 experiments for the first time.
This product contains all the materials essential for CRISPR-Cas9 experiments, including sgRNA, Cas9, Mutation Detection kit, etc.

Figure 1. sgRNA expression vector (dRGEN-U6-sgRNA)	Figure 2. Cas9 expression vector (pRGEN-Cas9-CMV/T7)

Both sgRNA and Cas9 will be given in the form of a plasmid, and three types of sgRNA will be also provided.
This kit needs pre-designed sgRNA sequences. If they are not ready yet, you may try BIONEER’s Custom Design Service to quickly prepare what you need.

• The sgRNA is cloned into a sgRNA expression vector (Figure 1).
• pRGEN-Cas9-CMV/T7 vector is provided in the plasmid form (Figure 2).
• The positive control will be provided. You can select one from the following: EGFP, CCR5, HPRT1.
• Double-strand breaks (DSBs) will be formed with the optimized genome editing solution.
• The genome editing results can be checked easily with the Mutation Detection kit by using T7E1.

√ We recommend these products to the following:

• Those who are just starting CRISPR-Cas9 experiments
• Those who wish to start CRISPR-Cas9 experiments, but are unsure which products to use
• Those who plan CRISPR-Cas9 experiments using plasmid forms

Features and Benefits

• Easy-to-use
  

  Perform all the steps of CRISPR-Cas9 experiments using only a single kit, having 3 types of sgRNA for sgRNA screening

• Convenience
  

  Easily perform validation tests with the Mutation Detection kit

• AccuTool ™ Safe Harbor Knock-In Kit

Overview

Safe Harbor is a genomic site allowing integration of the gene of interest into a specific region so that the novel inserted genes can be stably expressed.
The Safe Harbor Knock-in kit contains a gRNA expression plasmid optimized for targeting the safe harbor site and an HDR donor vector for performing the knock-in with high efficiency. With this, this product can be used to insert genes for a stable expression.

Knock-in have risks of mutation and gene silencing caused by random integration within the host genome.
Therefore, the new approach has been recently proposed to deliver the gene of interest to the ‘Safe Harbor’ site in the host genome (Figure 3).
The gene of interest can be expressed stably and consistently by inserting the gene of interest in the human AAVS1 genome site or the mouse Rosa26 genome site.

AccuTool™ Safe Harbor Knock-in kit contains gRNA expression vectors optimized for targeting human or mouse genomic safe harbor sites and an empty donor vector designed to effectively perform a target gene knock-in.

Figure 3. CRISPR-Cas9-based Knock-In mechanism
The donor vector containing the gene of interest, selection marker, or any other genetic elements is integrated into safe harbor site through the homologous recombination.

This product requires a donor vector cloned with the gene of interest to perform a knock-in.
If you have any difficulties preparing your donor template, try using BIONEER’s Gene Synthesis & Cloning Service.

AccuTool™ Safe Harbor Knock-In kit is divided into two products: AccuTool™ AAVS1 donor kit and AccuTool™ Rosa26 donor kit.
Depending on the kit, it contains an AAVS1_donor vector or a Rosa26_donor vector for cloning the gene of interest. It can be used with Cas9 expression vectors to deliver them to the host cell.

Those can then create DSBs in either the AAVS1 or Rosa26 site to activate HDR pathway, a natural DNA repair mechanism. It is used for the integration of donor DNA into the genomic safe harbor site. Furthermore, this kit contains primers to check the successful integration with PCR.

Features and Benefits

• Stability
  

  Genes can be expressed stably without any alternations using the genomic safe harbor site.

• Accuracy
  

  This kit enables site-specific gene integration with minimized off-targets.

Application

• These kits can be applied to clinical studies, gene functional studies, and genetic testing/analysis.