The emergence of the CRISPR-Cas9 system has triggered a technical revolution in mammalian genome editing. Compared to traditional gene-targeting strategies, CRISPR-Cas9 technology offers a more efficient and cost-effective approach for generating genetically modified animal models. However, off-target cleavage in CRISPR-mediated genome editing is a major concern in the analysis of phenotypes as well as the selection of therapeutic targets. 
Whole genome sequencing (WGS) is a straightforward approach for an unbiased survey of the full genome, which can be used to identify off-target nuclease activity  .
Genome-wide Off-target Detection Advantages
★ Unbiased, with the capability to screen out all potential off-target sites. 
★ The greatest sensitivity and precision for SNP/InDel detection. 
★ No cellular model restriction. 
★ Simple experimental designs. 
- Sample PreparationSample QC
- Library ConstructionLibrary QC
- SequencingSequencing QC
- Raw Data OutputData QC
- Bioinformatics AnalysisDelivery QC
 Dong Y, Li H, Zhao L. et al. Genome-Wide Off-Target Analysis in CRISPR-Cas9 Modifified Mice and Their Offspring. G3 (Bethesda).;9(11):3645-3651(2019).
 Atkins A, Chung CH, Allen AG, et al. Off-Target Analysis in Gene Editing and Applications for Clinical Translation of CRISPR/Cas9 in HIV-1 Therapy. Front Genome Ed.;3:673022(2021).
 Chen S, Yao Y, Zhang Y, Fan G. CRISPR system: Discovery, development and off-target detection. Cell Signal. 70:109577(2020).
 Foox, J., Tighe, S.W., Nicolet, C.M. et al. Performance assessment of DNA sequencing platforms in the ABRF Next-Generation Sequencing Study. Nat Biotechnol 39, 1129–1140 (2021).
CRISPR Genome-Wide Off-Target Analysis Specification
Sequencing Service Specification
- BGI Genome-wide Off-target Detection Services are performed with the DNBSEQ™ platform and standard bioinformatics pipeline for superior data quality and analysis results.
Sample Preparation and Services
- Library preparation (DNBSEQ™ platform).
- Human/Animal/Plant samples accepted.
- Appropriate sequencing strategies are recommended according to different data.
- Typical 31 working days from sample QC acceptance to data analysis report availability
- Expedited services are available; contact your local BGI specialist for details
|PCR||≥200ng (Recommend ≥400ng)||≥8ng/μL|
|PCR-free||≥1μg (Recommend ≥2μg)||≥12.5ng/μL|
- Assessment of sequencing reads
- Alignment of reads to reference genome, depth and coverage assessment
- SNP/InDel calling
- Specific variants detection (provide control sample)
- sgRNA homologous region analysis
- Editing efficiency analysis
- Off-target detection and statistics
- Off-target sites annotation
- Visualization of Analysis Results
- Further customization of bioinformatics analysis to suit your unique project is available: Please contact your BGI technical representative.