Overview

Off-target effects occur when small nucleic acid drugs bind to non-target mRNAs in addition to the intended target, potentially compromising both treatment safety and efficacy. GenScript combines RNA sequencing (RNA-seq) with advanced bioinformatics to detect transcriptional changes and pinpoint off-target activity- ensuring you choose the safest, most effective drug candidates.

Advantages

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Predictive Modeling

Predicts off-target effects to improve screening efficiency, saving both time and costs

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Reliable Detection

Optimized sample prep and proprietary NGS workflows deliver accurate, reproducible results

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Actionable Analytics

Includes raw data, data QC, and multidimensional differential expression reports for confident decision-making

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Comprehensive Service Workflow

Off-Target Prediction

Off-Target Prediction

  • Predict off-target effects based on candidate sequences and target genes.
  • Select candidates with lower off-target potential for further testing.
RNA-seq Off-Target Detection

RNA-seq Off-Target Detection

  • Transfect cells with siRNA samples
  • Extract total RNA
  • Select mRNA and prepare libraries
  • Perform NGS
  • Conduct data analysis and visualization
Experimental Validation

Experimental Validation (Optional)

Assess the impact of candidate sequences on the transcription of different genes at the mRNA level using RT-PCR

Off-Target Optimization

Off-Target Optimization (Optional)

  • Seven proprietary modification strategies designed to reduce off-target effects.
  • Prediction of knockdown efficiency to guide optimal selection.

Service Details

Item Service Content
Applicable Scope siRNA, ASO, miRNA and other small nucleic acid drugs
Experimental Plan
  • Bioinformatics tools for off-target effect evaluation (miRNA off-targeting*, Homologous off-targeting**, etc.)
  • RNA-seq detection of differentially expressed genes (DEGs), down-regulated off-targeting***, etc.
Report
  • Bioinformatics evaluation report
  • Raw sequencing data (FASTQ)
  • Data QC analysis
  • DEG analysis
  • Visualization (heatmaps, volcano plots, GO enrichment, KEGG enrichment analysis, etc.)
Timeline Starting from 2 weeks
Additional Services (Optional)
  • Off-target validation (RT-PCR)
  • Modification optimization of candidate sequences for reduced off-target effects

* miRNA Off-Targeting: siRNA seed region similarity to target gene leads to regulation of non-target genes via a mechanism similar to miRNA.
** Homologous Off-Targeting: siRNA sequence shares homology with non-target genes, causing unintended interference.
*** Down-Regulated Off-Targeting: Non-specific mechanisms reduce expression of non-target genes.

Case Studies

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