When the CRISPR/Cas9 system enters the body, the primers specifically identify the gene to be edited and its sequence position, directing Cas9 to perform random cleavage of the first few bases of its recognition site (PAM sequence). However, this recognition is not absolutely specific and may recognize other similar sequences in the genome and erroneous gene editing may occur. This is a pressing problem and bottleneck in the gene editing process. Errors in editing can lead to off-target effects and as a result can lead to mutations in other non-target genes. Artificial mutations in non-target genes can be accompanied by the development of various serious diseases with serious consequences. Therefore, off-target detection is very important in the gene editing process. Moreover, how to effectively identify and avoid the effects of off-target effects has been a key research direction.
Based on this, CD Genomics provides you with high-throughput NGS-based tools to identify and detect off-target sites and help you optimize your gene editing strategy to reduce the occurrence of off-target effects. Gene editing is only meaningful if the target gene is correctly targeted, and detection of off-target effects is the most important step in the development of a gene editing strategy.
Our SureGuide OnTarget Platform is a powerful tool for assessing off-target effects. The principle is to use a short double-stranded oligonucleotide tag to mark CRISPR-Cas-induced breaks (on-target and off-target), followed by high-throughput sequencing of the gene region where the tag is located, and finally bioinformatics analysis to determine the location of the off-target mutation and the mutation frequency.
CD Genomics has been committed to making a difference in your off-target detection services through high-throughput sequencing platforms to support CRISPR optimization services during gene editing. The importance of off-target detection directly determines the success of gene editing. Our CRISPR off-target detection service process is shown in the figure below, and the whole process is operated by a professional team of experts under strict QC standards.
01. Sequencing sample requirements
Cell lines after gene editing (frozen or in culture dishes)
02. Library construction and NGS
NGS-based Illumina / MGI platform
03. Bioinformatics Analysis
CRISPR off-target assays are an important indicator for gene editing safety validation. CD Genomics provides NGS-based off-target assay sequencing services to enable efficient optimization of drug development and biotech company pipelines. In gene editing, CRISPR off-target assays are used for off-target prediction and assessment, off-target rate optimization, validation of CRISPR gene editing, gene editing safety assessment, and driving the drug screening process.
The application of CRISPR screening holds substantial prospects in unravelling genes responsible for various physiological phenomena, including drug resistance and sensitivity, thereby establishing itself as an indispensable tool within biomedical and pharmaceutical research. Here at CD Genomics, we utilize Next-Generation Sequencing (NGS) as part of our high-throughput strategies to bolster your CRISPR screening process. This strategy ensures a combination of efficacy and superior quality in the screening and subsequent analysis of your targeted genes.
CRISPR/Cas9, despite its renown as an avant-garde gene editing tool, is not wholly exempt from challenges such as off-target mutations, erroneous allele modifications, or unnecessary redundant edits. Accordingly, it becomes pivotal to verify the accuracy of genome editing post the implementation of CRISPR/Cas9 experiments. By harnessing the power of Next-Generation Sequencing (NGS)- anchored high-throughput methodologies, CD Genomics stands in a position to offer efficient, precise validation services for your gene editing outcomes.
CD Genomics is committed to combining professional project protocol guidance and standardized analysis processes to ensure that your project is performed accurately and quickly. If you are interested in CD Genomics' CRISPR off-target discovery in the field of genome editing and engineering, please feel free to contact us now.
CD Genomics is transforming biomedical potential into precision insights through seamless sequencing and advanced bioinformatics.
