Sinteza saRNA po meri
Samoojačevalna mRNA (saRNA), znana tudi kot replikonska RNA, kodira zaporedja mRNA in vitro transkripcije (IVT) in gene virusne replikaze, pridobljene iz alfavirusa ali flavivirusa. Geni virusne replikaze omogočajo samopomnoževanje mRNA, kar ima za posledico povečano izražanje beljakovin in najmanjši zahtevani odmerek RNA v primerjavi z nepomnoženimi mRNA. Drugi vidik je, da je saRNA razmeroma velika molekula (več kot 14 kb).
Yaohai Bio-Pharma je vzpostavil nabor tehnologij za sintezo mRNA za zagotavljanje mRNA brez pomnoževanja in mRNA s samopomnoževanjem (1000 nt~14000 nt), kot so zasnova in optimizacija zaporedja, IVT, čiščenje, liofilizacija in inkapsulacija lipidnih nanodelcev (LNP). . Vsi izdelki so izdani v skladu s strogimi standardi nadzora kakovosti.
Fig.1 Structural Features of IVT mRNAs: Conventional, Circular and Self-amplifying mRNA.
Sequence Features of Self-amplifying mRNA
The self-amplifying RNA comprises four non-structural proteins (designated as nsP1, nsP2, nsP3, and nsP4), which are derived from alphaviruses, Venezuelan equine encephalitis virus (VEEV). Among these, nsP1 exhibits dual enzymatic activities, namely GTase and N7MTase, whereas nsP2 possesses RTPase activity, pivotal for capping the IVT mRNA to generate Cap 0 structure. Additionally, nsP2 serves as a protease and helicase, facilitating the intricate processing of the entire nsP complex. The precise function of nsP3 remains elusive, yet it engages in interactions with diverse host cell proteins, thereby contributing to the attenuation of the antiviral response mounted by the host.
An alphavirus subgenomic promoter (SGP) region is located before the gene of interest (GOI). The SGP element facilitates the initiation of GOI transcription by bypassing the reading of the sequence encoding viral nsP proteins.
Fig.2 Synthesis and cell assays of eGFP saRNA of Yaohai Bio-Pharma RNASci Platform.
Custom Solutions of saRNA
Protokol sinteze saRNA je podoben mRNA: Sinteza mRNA po meri
- Oblikovanje zaporedja
- Priprava plazmidne šablone
- in vitro transkripcija (IVT)
- Encimsko omejevanje mRNA
- Omejitev kotranskripcije mRNA
- Čiščenje mRNA
- Liofilizacija mRNA
- Enkapsulacija LNP
- Analiza in testiranje
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