About AKT1
The AKT1 gene (AKT Serine/Threonine Kinase 1) on chromosome 14q32 The E17K mutation in the pleckstrin homology domain is a recurrent activating mutation found in breast, ovarian, and colorectal cancers. Key targets include Exons 2–6 (PH domain, kinase domain) for comprehensive mutation screening.
Mutations: E17K, Q79K | GC content: 52% | Target exons: Exons 2–6 (PH domain, kinase domain)
AKT1 Primer Design Challenges
- Hotspot mutation E17K: The recurrent E17K mutation in exon 2 requires allele-specific primer design for mutant detection
- Low sequence complexity: Intronic regions around exon 4 contain repetitive elements complicating unique primer placement
- Pseudogene interference: Processed pseudogene AKT1P on chromosome 14 shares>90% exon homology requiring careful specificity checks
Recommended Primer Design Parameters for AKT1
| 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 AKT1
| Target Region | Forward Primer (5′→3′) | Reverse Primer (5′→3′) | Amplicon |
|---|---|---|---|
| Exon 2 (E17K hotspot) | 5′-GGTGCCATTGAGAAGGACGT-3′ | 5′-ACAGGTGGTCCACATCCTCT-3′ | 178 bp |
| Exon 4 (kinase domain) | 5′-TCTGTCATGGAGTACGCCAA-3′ | 5′-TGTGTCCTGGTCCTGGTTCT-3′ | 203 bp |
Key SNPs to Avoid in Primer Binding Sites
When designing AKT1 primers, avoid these clinically significant variants:
- rs121434592 (E17K) — PH domain hotspot in exon 2
- rs1130233 (Q79K) — Common polymorphism near the kinase domain
Clinical Validation Required
All AKT1 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 AKT1 Primers Now
Pre-configured with AKT1-specific parameters. Use our primer design, Tm calculator, and GC content tools for optimal results.