體外轉錄線性mRNA合成服務
IVT linear mRNA Synthesis Service
有鑑於 mRNA 的安全性 (無基因嵌入風險) 及高表達效率 (細胞質內快速轉譯),使得 mRNA 於近年來被廣泛應用於各式疫苗開發、癌症免疫療法、細胞治療、基因編輯等面向。近幾年新冠疫情衝擊全球,藉由接種可轉譯出 SARS-CoV-2病毒棘蛋白的 mRNA疫苗,能夠讓人體及早獲得保護力,降低感染及重症的風險。因此高品質的 mRNA 將大幅加速 mRNA 療法及相關應用的發展。
◆ 提供 RUO 與 Pre-clinical 等級的高純度 IVT mRNA合成服務
◆ 擁有一站式解決方案,從密碼子優化、基因合成與選殖、質體製備、mRNA 生合成至 LNP 封裝,均能一次滿足您的需求
◆ mRNA 共轉錄式加帽效率 (co-transcriptional capping rate) 高於 93%
◆ 均質的 Poly (A) tail,能強化 mRNA 表達效率,可保證提供 100A Tail
◆ 多樣化 mRNA 純化方式和QC策略,可供彈性選擇
◆ 擁有豐富經驗的生產團隊和優化平台,能幫您省下不少設計和製作 mRNA 的時間和精力
IVT mRNA One-stop Solution (From gene synthesis to LNP Encapsulation)
Gene Synthesis (Codon optimization) & Cloning |
⮕ |
Plasmid Synthesis & Linearization |
⮕ | mRNA Synthesis & Purification | ⮕ | Lipid Nanoparticles Encapsulation |
IVT mRNA Synthesis Workflow
◆ 合成長度:100 bp – 20 kb
◆ 合成量:100 ug – 200 mg
◆ Poly (A) Tail 選項:100A Tail (推薦)、Custom tail ≤ 120A、No Tail
5' Cap 選項:Cap0、Cap1 (Co-Capped or enzymatic)(推薦)、Cap2 (Co-Capped)、Uncapped
◆ Modified NTP選項:N1-Me-Ψ (推薦)、5moU、5meC & Ψ、Unmodified
◆ 緩衝液選項:1 mM Sodium Citrate (pH 6.5)(預設)、TE (10 mM Tris, 0.1mM EDTA, pH 7.0)、RNase-free water
純化方式
(1) RUO 等級:Silica membrane-based or LiCl precipitation method
(2) Pre-clinical 等級:HPLC-based Oligo dT Purification
(3) 升級純化方式:dsRNA Removal
QC 項目
Agarose gel electrophoresis、pH、A260/280 ratio、Semi-quantitative Endotoxin test
加價版QC服務
包含毛細管電泳、LC-MS (檢測capping效率)、蛋白殘留量測試 (≤ 1%)、質體殘留測試 (≤ 0.05%)、Slot-blot (檢測dsRNA殘留)、生菌試驗 (No Growth after 48 hrs) 等。
Cap 0 | Cap 1 | Cap 2 | |
Structure | N7-methyl guanosine | N7-methyl guanosine + 2'O-methylation on first nucleotide | N7-methyl guanosine + 2'O-methylation on first and second nucleotide |
Prevalence | Plants and RNA viruses | Vertebrates | Vertebrates |
Functions | Protection from degradation mRNA translation efficiency | Protection from degradation mRNA translation efficiency Distinguish self mRNA from viral mRNA | Protection from degradation mRNA translation efficiency Distinguish self mRNA from viral mRNA Reduce activation of innate |
*Diagram of Cap structures above was retrieved with some modifications.7
◆ IVT linear mRNA in desired buffer
◆ COA report
上述屬客製化服務,如想動手做 IVT mRNA,可參考下列品牌的相關產品
TAKARA – Cloning & mRNA synthesis & post-transcriptional capping