Documentation
Getting Started
T-GUIDE identifies efficient guide RNAs for TnpB and CRISPR-Cas systems across 9 model organisms. Follow these steps to design your guides:
Select Genome
Choose from Human (hg38), Mouse (mm39), Rat, Chicken, Cow, Pig, Zebrafish, C. elegans, or Drosophila.
Choose Nuclease
Select SpCas9 (NGG PAM) or one of 50+ TnpB variants with their specific PAM requirements.
Enter Target
Enter a gene symbol (e.g., BRCA1) or paste a custom DNA sequence.
Analyze Results
Review guides in the table, visualize on the genome browser, and run off-target analysis.
Gene Search Mode
Enter a gene symbol to automatically fetch the canonical transcript and design guides across all exons:
- Gene lookup uses Ensembl's REST API
- Canonical transcript is selected automatically
- Exon/intron structure is visualized in the browser
- Guides are classified as Exon or Intron based on location
Sequence Mode
Paste raw DNA or FASTA format to find guides in custom sequences:
- Supports plain sequence or FASTA with header
- GC content and length calculated in real-time
- Useful for targeting non-coding regions or novel sequences
Genome Browser
Interactive SVG-based visualization showing:
- CDS: Coding sequences (Tall boxes)
- UTR: Untranslated regions (Thin boxes)
- Forward (+): Guides on plus strand
- Reverse (-): Guides on minus strand
Off-Target Analysis
Powered by Cas-OFFinder, our off-target analysis searches for potential mismatches:
- MM0: Exact matches (should be 1 for the target)
- MM1-3: Sites with 1-3 mismatches
- Click individual guides to view detailed off-target sites
- Use "Analyze All" for batch processing