Overview of CRISPR validation sequencing

CRISPR/Cas9 system has been continuously improved and has gradually become a powerful tool suitable for gene editing in all kinds of organisms. Whether it is single base editing, small fragment deletion, mutation at the DNA level, transcriptional regulation of RNA, RNA interference at the RNA level, or epigenetic-based regulation of DNA methylation, it has brought great convenience to researchers. sgRNA libraries of CRISPR/Cas9 combined with high-throughput screening technology have a wide range of applications, both genome-wide screening to discover new targets, and also to quickly and accurately find genes or gene clusters associated with a certain phenotype. During gene editing involving CRISPR/Cas9, there are two methods to repair the sheared duplex, including non-homologous end joining (NHEJ) or homologous directed repair (HDR).

Figure 1. Schematic diagram of the repair methods of the sheared strand during gene editing.

However, although the CRISPR/Cas9 system has become a hot gene editing tool, there are still problems such as missed edits, wrong edits into alleles or duplicate edits. To avoid these problems, it is important to validate genome editing after CRISPR/Cas9 experiments. With NGS-based high-throughput tools, CD Genomics can provide you with efficient and accurate gene editing result validation services. The entire process involves our experienced experts and is maintained under strict QC standards. We provide you with an end-to-end solution, you only need to provide the samples to be tested as specified. Ultimately you will receive a comprehensive and detailed analysis of the results of CRISPR validation sequencing.

The workflow of CRISPR validation sequencing service

CD Genomics is committed to providing NGS-based CRISPR validation sequencing services to help analyze target activity and predict editing effects. CRISPR validation is an important guide for gene editing and target screening, and the sequencing process is shown in the figure below. The whole process follows 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

• Reference alignment
• Visual annotation of target gene loci
• Target activity analysis
• CRISPR feasibility assessment analysis
• CRISPR validation result report

Advantages of our CRISPR validation sequencing

• NGS-based high-throughput technology platform ensures sequencing depth and high sensitivity
• One-stop sequencing analysis, from the sample to be tested directly to the analysis report, greatly improves efficiency
• Effectively simplifies the complex CRISPR validation process and enables high-throughput validated sequencing
• CRISPR validation results reporting ensures comprehensive and detailed reporting, while our experienced experts provide advice and consultation for your CRISPR improvement and design

Applications in genome editing and engineering

CD Genomics provides NGS-based high-throughput CRISPR validation methods that improve efficiency while ensuring the accuracy of the assay. The purpose of CRISPR validation sequencing is to analyze gene editing results and detect target activity, mainly to guide the design, optimization and feasibility study of CRISPR/Cas9, which is widely used in various disease pathogenesis studies, target It is widely used in various disease mechanisms, target activity screening and drug discovery.

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 validation sequencing in the field of genome editing and engineering, please feel free to contact us now.