About CARD11
The CARD11 gene (Caspase Recruitment Domain Family Member 11) on chromosome 7p22 Gain-of-function mutations in the coiled-coil domain (exons 3–10) drive constitutive NF-κB activation in diffuse large B-cell lymphoma. Key targets include Exons 3–10 (coiled-coil domain) for comprehensive mutation screening.
Mutations: L232LI, G123S, D230N | GC content: 51% | Target exons: Exons 3–10 (coiled-coil domain)
CARD11 Primer Design Challenges
- Coiled-coil domain mutations: Hotspot L232LI insertion in exon 6 requires careful amplicon placement around the repeat region
- Alternative splicing: Multiple CARD11 isoforms with alternative exon usage require transcript-aware primer design
- Moderate GC content (51%): Balanced GC provides favorable thermodynamics but requires uniform Tm across target regions
Recommended Primer Design Parameters for CARD11
| Parameter | Standard Exons | Optimized Regions |
|---|---|---|
| Primer length | 20–22 nt | 22–25 nt |
| GC content | 45–55% | 50–60% |
| Tm | 58–62°C | 60–65°C |
| Amplicon size | 150–300 bp | 180–350 bp |
| Annealing temp | 58–60°C | 60–64°C (touchdown) |
| PCR additive | Standard | Optional 3–5% DMSO for problematic regions |
Recommended Primer Sequences for CARD11
| Target Region | Forward Primer (5′→3′) | Reverse Primer (5′→3′) | Amplicon |
|---|---|---|---|
| Exon 6 (coiled-coil) | 5′-GAACGAGGTGCTGCTGCTAA-3′ | 5′-CAGCAGCTCCACAGACACTT-3′ | 183 bp |
| Exon 4 (CARD domain) | 5′-AGCCCTGTGGACTTCCTCTT-3′ | 5′-ACCTGGCGTGGTTAAAGTCC-3′ | 156 bp |
Key SNPs to Avoid in Primer Binding Sites
When designing CARD11 primers, avoid these clinically significant variants:
- rs201569880 (L232LI) — Exon 6 insertion in DLBCL
- rs587782115 (G123S) — Exon 4 CARD domain variant
Clinical Validation Required
All CARD11 primers designed with VigyanLLM are for research use only. Clinical diagnostic applications require additional wet-lab validation, Sanger sequencing confirmation, and regulatory approval before patient use.
Design CARD11 Primers Now
Pre-configured with CARD11-specific parameters. Use our primer design, Tm calculator, and GC content tools for optimal results.