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Protocols

protocols
Protocol

CRISPR/Cas9 + AAV-Mediated Intra-Embryonic Gene Knocking Protocol in Mice

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: This experiment describes the method of transferring plasmid DNA molecules into <em>E. coli</em> and how to screen cell clones with plasmid DNA, i.e. transformants.

protocols
Protocol

Preparation of dsRNA by In Vitro Transcription Protocol

Experiment Tags: dsRNA Synthesis, <em>In Vitro</em> Preparation

Experiment Summary: In this protocol, we describe methods for the production of stable luciferase expressing tumor cell lines by lentiviral transduction.

protocols
Protocol

Protocol for Selecting Vaccinia Virus Recombinants Using CRISPR/Cas9

Experiment Tags: Recombinant Virus, Vaccinia Virus, VACV (VACV), CRISPR/Cas9 Selection, Virotherapy, Vaccine Development

Experiment Summary: This protocol describes a new method, the AAV-Cpf1 KIKO system, using a combination of viral and non-viral approaches to generate a stable CAR-T with homology-directed repair (HDR) knock-in and immune checkpoint knockout at high efficiency in one step.

protocols
Protocol

Protocol for Transient yet Robust Expression of Proteins in the Mouse Liver via Intravenous Injection of Lipid Nanoparticle-encapsulated Nucleoside-modified mRNA

Experiment Tags: Nucleoside-Modified mRNA, Lipid Nanoparticle, Liver Regeneration, Protein Expression, Retro-orbital Injection

Experiment Summary: The protocol introduces the definition of cell immortalization and commonly used viral transfection methods, and describes in detail the steps to generate immortalized cells using prefabricated cell immortalization lentiviral particles.

protocols
Protocol

Protocols for In vitro Transcription (IVT) and tRNA Binding Assay

Experiment Tags: T-box (T-box), EMSA (EMSA), RNA-RNA Interaction, TRNA (tRNA), In Vitro Transcription

Experiment Summary: This protocol discusses the production, purification, and use of replication-deficient retroviral vectors based on Moloney murine leukemia virus and lentivirus. It also describes the injection of a retroviral vector into the dentate gyrus of young adult mice to fluorescently label live murine brain tissue.