Journal: Viruses

Article Title: A mRNA Vaccine for Crimean–Congo Hemorrhagic Fever Virus Expressing Non-Fusion GnGc Using NSm Linker Elicits Unexpected Immune Responses in Mice

doi: 10.3390/v16030378

Figure Lengend Snippet: Genomic structure of mRNA template. ( A ) Corresponding Cap1-mRNA were synthesized by IVT and capping using constructed pGEM-3Zf (+) mt including target antigen as templates; 5′UTR was α-globin, 3′UTR was mtRNA1-AES, and poly A tail included 120 A. The short arrows indicate enzyme digestion. ( B ) Target antigens were eGFP, Luc, Gn, GnHis, Gc, GcHis, GnNSmGc, and GnNSmGcHis. The red box indicates immunogen genes encoding CCHFV glycoprotein that were only codon-optimized for humans. The dotted line indicates that GnNSmGc and GnNSmGcHis could be digested to Gn, NSm, and Gc/GcHis by SKI-1/S1P and SKI-1/S1P-like enzymes. The short arrows indicate sites for enzyme digestion.

Article Snippet: mRNA was produced and purified by T7 High Yield RNA Synthesis Kit (E2040S, NEB, Beijing, China), Vaccinia Capping System (M2080S, NEB, Beijing, China), pseudouridine (ψ) (N-1019, TriLink, Shenzhen, China), mRNA Cap 2′-O-Methyltransferase (M0366S, NEB, Beijing, China), DNase I (M0303S, NEB, Beijing, China), and Monarch ® RNA Cleanup Kit (T2050L, NEB, Beijing, China), as follows: (a) Linearize 8 μg of pGEM-3Zf (+) mt-eGFP/Luc/Gn/GnHis/Gc/GcHis/GnNSmGc/GnNSmGcHis plasmid with 4 μL NsiI-HF enzyme (R3127, NEB, Beijing, China) overnight at 37 °C. (b) Verify above plasmids for complete linearization by gel electrophoresis, before purification with Cycle-Pure Kit (D6492, Omega, Beijing, China). (c) Formulate reaction system ( ) in tubes and perform in vitro transcription (IVT) for 10 h at 37 °C.

Techniques: Synthesized, Construct

Journal: iScience

Article Title: Sirt6 regulates the proliferation of neural precursor cells and cortical neurogenesis in mice

doi: 10.1016/j.isci.2023.108706

Figure Lengend Snippet:

Article Snippet: Plasmid: pGEM-3zf (+) , Promega , Cat#P2271.

Techniques: Recombinant, Imaging, Western Blot, Sequencing, Plasmid Preparation, Software