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Creative Biogene CRISPR/Cas9 Platform provides a GUIDE-Seq service to assess off-target effect of CRISPR/Cas9. At CRISPR/Cas9 PlatformCB, all projects are performed by skillful staff under the guide of experienced scientists. We are dedicated to assisting you and paving the way for your successful research with reliable services and products.
Since CRISPR/Cas9 technology was applied for genome engineering, it brings a revolutionary development of genome-editing technology. CRISPR system makes it easy and efficient to make targets cuts to genome. However, it may induce cleave off-target DNA targets at low frequencies when applied for genome editing. To overcome this limitation, various methods have developed to detect these unexpected cleavages by scientists. Genome-wide Unbiased Identifications of DSBs Evaluated by Sequencing (GUIDE-Seq) is one of cell-based assays for off-target sites detection. This assay is an approach for global detection of DNA double-stranded breaks introduced by RGNs (CRISPR RNA-guided nucleases) and potentially other nucleases. It is highly sensitive and can detect off-target sites that occur at a frequency of 0.1% in a cell population. GUIDE-Seq is a recommended method to detect the putative off-target cleavage sites.
Fig 1. Schematic overview of GUIDE-Seq method1
GUIDE-Seq consists of two stages. At the first stage, RGNs are transfected into living cells and then induce double-stranded breaks. Then small, double-stranded oligonucleotides (dsODN) are inserted to genome by NHEJ pathway and tagged these breaks. At the second stage, these break sites are identified by unbiased amplification and next-generation sequencing. GUIDE-Seq identifies off-target sites genome-wide, including exons, introns and intergenic regions. And it is adapted to assess the genome-wide specificities of nucleases, including dimeric ZFNs, TALENs and CRISPR RNA-guided nucleases.
CRISPR/Cas9 PlatformCB provides a complete GUIDE-Seq service, including sample preparation, sequencing and data analysis. Once a project finished, we will provide a detailed reporter with experiment results and raw data to customers. Custom service is welcome as well.