CRISPR gRNA Design
AI-powered gRNA design for target gene editing
High-Performance Client-Side CRISPR Guide RNA Design & Analysis Platform
Top 15 gRNA Candidates
| Rank | Position | Guide Sequence | PAM | GC% | On-target | Off-targets | Warnings |
|---|---|---|---|---|---|---|---|
| 1 | 75 + | GCCTCAGGCTGAGCTGGGGG | CGG | 75% | 0.2773 | - | |
| 2 | 149 + | GGGTCATTGGGAGGGGGGGG | CGG | 75% | 0.2024 | - | |
| 3 | 28 + | GGGGACCTGGAGTCGTGGGG | TGG | 75% | 0.1092 | - | |
| 4 | 56 - | TCCGCCCCCAGCTCAGCCTG | AGG | 75% | 0.1035 | - | |
| 5 | 87 - | ACCCCTTGTCCCCTCCCCCC | CGG | 75% | 0.102 | - | |
| 6 | 119 + | GGACAAGGGGTGACAGGCTG | CGG | 65% | 0.0978 | - | - |
| 7 | 140 + | GGGGCTACGGGGTCATTGGG | AGG | 70% | 0.0925 | - | - |
| 8 | 146 + | ACGGGGTCATTGGGAGGGGG | GGG | 70% | 0.0803 | - | - |
| 9 | 113 + | GGGAGGGGACAAGGGGTGAC | AGG | 70% | 0.057 | - | - |
| 10 | 78 + | TCAGGCTGAGCTGGGGGCGG | AGG | 75% | 0.0544 | - | |
| 11 | 82 + | GCTGAGCTGGGGGCGGAGGG | TGG | 80% | 0.0534 | - | |
| 12 | 185 + | GGGAGACAGCTGCGGGGCTA | CGG | 70% | 0.0476 | - | - |
| 13 | 143 + | GCTACGGGGTCATTGGGAGG | GGG | 65% | 0.0451 | - | - |
| 14 | 60 + | TCAGCTCTGGGGCTCGCCTC | AGG | 70% | 0.0444 | - | - |
| 15 | 158 + | GGAGGGGGGGGCGGATGGGG | AGG | 85% | 0.0379 | - |
Molecular Analysis of Selected gRNA
Select a gRNA candidate from the table to inspect its molecular structure, score breakdown, off-target map, and cloning oligos.
Tool Guide
Definition
CRISPR gRNA design tool — Input target DNA sequence for 3-variant PAM scanning (SpCas9/SaCas9/AsCas12a) + Doench 2014 Rule Set 1 on-target scoring + self-off-target search + 4 biochemical filters + automatic cloning oligo design. 100% client-side.
Purpose
(1) Select first-pass gRNA candidates for knockout cell line construction (2) Exon-specific targeting (prefer early exons) (3) Filter candidates with minimal off-target hits (4) Design multi-gRNA libraries for saturating mutagenesis
How to Use
① Enter target DNA sequence (FASTA supported, or use TP53/BRCA1 sample buttons) ② Select Cas variant (SpCas9 NGG / SaCas9 NNGRRT / AsCas12a TTTV) ③ Choose cloning vector (pSpCas9-BbsI / pX601-BsmBI / None) ④ Adjust GC% filter range (default 40-70%) ⑤ Top 15 candidates auto-generated → click for detailed analysis: • DNA-gRNA R-Loop hybridization visualization • Doench 2014 position-specific score breakdown • Self-off-target map within input sequence • Cloning oligos (Top/Bottom) + copy buttons
Examples
Example) Design gRNA to KO TP53 exon 4 in human → Input: gene = TP53, genome = GRCh38 → Output: 5'-GAGCGCTGCTCAGATAGCGA-3' (PAM: TGG, exon 4) • On-target: 0.78 (high) • Off-target: 0 perfect match, 2 sites with 3-mismatch → Synthesize → clone into lentiCRISPRv2 → HEK293 transduction