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Experiment Tags: T-box (T-box), EMSA (EMSA), RNA-RNA Interaction, TRNA (tRNA), In Vitro Transcription
Experiment Summary: This protocol describes the coupling of (i) “live” in vitro RNA transcription with (ii) binding by a radiolabeled, pre-formed tRNA followed by native gel electrophoresis and phosphorimager scan to visualize the complex.
Experiment Tags: Respiratory Syncytial Virus, Respiratory Infection, Virus Detection
Experiment Summary: In this protocol, we discuss in detail how to infect BALB/c mice with RSV and how to detect the RSV genome in mouse lungs using reverse transcription quantitative PCR (RT-qPCR).
Experiment Tags: Zika Virus, NS4B (NS4B), Double-Stranded RNA, Virus Titration, Immunofluorescence Microscopy, Plaque Assay
Experiment Summary: In this protocol, we describe basic experimental procedures to produce ZIKV stocks and to quantify their concentration in infectious virus particles as well as to image and study this pathogen within infected cells using confocal microscopy-based imaging.
Experiment Tags: Chikungunya, Enzyme-Linked Immunosorbent Assay (ELISA)
Experiment Summary: In this protocol, we accurately describe specific assays that can deterministically detect anti-chikungunya virus antibodies (Ab) in the tissues or sera of infected mice.
Experiment Tags: Nucleoside-Modified mRNA, Lipid Nanoparticle, Liver Regeneration, Protein Expression, Retro-orbital Injection
Experiment Summary: In this protocol, we describe in detail the necessary steps to deliver mRNA-LNP to the murine liver and, following intravenous injection of eGFP mRNA-LNP, verify transfection efficiency using flow cytometry and liver cell specificity using immunofluorescence analyses.
Experiment Tags: Recombinant Virus, Vaccinia Virus, VACV (VACV), CRISPR/Cas9 Selection, Virotherapy, Vaccine Development
Experiment Summary: In this protocol, we describe protocols for the generation and enrichment of recombinant vaccinia viruses using targeted Cas9 as a selection tool. This novel use of Cas9 is a simple addition to current homologous recombination-based methods that are widespread in the field, facilitating implementation in laboratories already working with poxviruses.
Experiment Tags: Subcellular Fractionation, RT-qPCR, Circular RNA, circRNA, Cytoplasmic circRNA, Nuclear circRNA, Low-Abundance circRNA
Experiment Summary: In this protocol, we present a protocol for the subcellular fractionation of cells to detect circRNA via RT-qPCR with divergent primers. Moreover, detailed steps for the generation of specific circRNAs-enriched cDNA included in this protocol will enhance the amplification and detection of low-abundance circRNAs.
Experiment Tags: dsRNA Synthesis, In Vitro Preparation
Experiment Summary: In this protocol, we describe a simple and widely used in vitro transcription reaction for the preparation of double-stranded RNA using PCR templates. The dsRNA prepared by this method can be used to induce RNA interference in certain cells or tissues.
Experiment Tags: CRISPR/Cas9 (CRISPR/ Cas9), Adeno-Associated Viral Vector, Trans-Zona Pellucida, Intra-Embryo Genome Editing, Ribonucleoprotein Electroporation, Large Fragment Knock-In
Experiment Summary: In this protocol, we demonstrate a detailed protocol for AAV-mediated intra-embryo knock-in in laboratory mice. This technique does not require either micromanipulation or germline competent pluripotent stem cells.
Experiment Tags: Biological Techniques, Recombinant, Lentivirus, Transduction
Experiment Summary: In this protocol, we describe the transduction of cultured cells with recombinant lentivirus expressing fluorescent markers.
