CRISPR Genome-Wide Off-Target Analysis

Description
Specification
RESOURCES

Service Description

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. [1] 

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 [2] . 

Genome-wide Off-target Detection Advantages

★ Unbiased, with the capability to screen out all potential off-target sites. [3] 

★ The greatest sensitivity and precision for SNP/InDel detection. [4] 

★ No cellular model restriction. [1] 

★ Simple experimental designs. [1] 

Project Workflow

We care for your samples from the start through to the result reporting. Highly experienced laboratory professionals follow strict quality procedures to ensure the integrity of your results.
  • Sample Preparation
    Sample QC
  • Library Construction
    Library QC
  • Sequencing
    Sequencing QC
  • Raw Data Output
    Data QC
  • Bioinformatics Analysis
    Delivery QC

References

[1] 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).

[2] 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).

[3] Chen S, Yao Y, Zhang Y, Fan G. CRISPR system: Discovery, development and off-target detection. Cell Signal. 70:109577(2020).

[4] 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

BGI Genome-wide Off-target Detection Services are performed on the DNBSEQ™ platform.

  • 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.

  • Turnaround Time

    • Typical 31 working days from sample QC acceptance to data analysis report availability
    • Expedited services are available; contact your local BGI specialist for details 

Sample Requirements

DNA Sample: Regular Samples
Library typeMassConcentration
PCR≥200ng (Recommend ≥400ng)≥8ng/μL
PCR-free≥1μg (Recommend ≥2μg)≥12.5ng/μL

Data Analysis

Besides data output, BGI offers a range of standard and customized bioinformatics pipelines for your project. Reports and output data files are delivered in these file formats: FASTQ, BAM, VCF, JSON and TXT.
STANDARD BIOINFORMATICS ANALYSIS
  • 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
CUSTOM ANALYSIS
  • Further customization of bioinformatics analysis to suit your unique project is available: Please contact your BGI technical representative.

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