Full Description
Synthetic mRNA is an attractive tool for mammalian cell reprogramming that can be used in basic research, as well as in clinical applications. Present mRNA in vitro synthesis is a rather simple procedure, which delivers a high yield of quality product. Various modifications may be introduced into the mRNA by changing the sequence of the DNA template, by modifying the reaction of transcription, or by post-transcriptional modification. mRNA, as a transfection agent, has several advantages over DNA, as mRNA expression is not dependent on nuclear entry and occurs directly in the cytosol. Synthetic Messenger RNA and Cell Metabolism Modulation: Methods and Protocols covers the typical main methods, such as mRNA synthesis, modifications, and delivery. Examples of cell reprogramming and analysis in the fields of immunotherapy and stem cell research are also included. Written in the successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and easily accessible, Synthetic Messenger RNA and Cell Metabolism Modulation: Methods and Protocols will be of interest to researchers, clinicians, and biotech companies interested in mRNA-mediated cell reprogramming.
Contents
Cell Engineering with Synthetic Messenger RNA.- In vitro Transcription of Long RNA Containing Modified Nucleosides.- HPLC Purification of in vitro Transcribed Long RNA.- Synthetic mRNAs with Superior Translation and Stability Properties.- Non-Viral, Cationic Lipid-Mediated Delivery of mRNA.- Light-Induced mRNA Transfection.- Engineering B Cells with mRNA.- Programming Human Dendritic Cells with mRNA.- Large Volume Flow Electroporation of mRNA: Clinical Scale Process.- Retrovirus-Based mRNA Transfer for Transient Cell Manipulation.- mRNA Delivery to Human Dendritic Cells by Recombinant.- Non-Viral RNA Transfection to Transiently Modify T Cells with Chimeric Antigen Receptors for Adoptive Therapy.- Natural Killer Cell Reprogramming with Chimeric Immune Receptors.- Reprogramming to Pluripotency and Differentiation of Cells with Synthetic mRNA.- Anti-Tumor Vaccination with Synthetic mRNA: Strategies for in vitro and in vivo Preclinical Studies.- Mannosylated and Histidylated LPR Technology for Vaccination with Tumor Antigen mRNA.- Analysis of Surviving-Specific T Cells in Breast Cancer Patients Using Human DCs Engineered with Survivin mRNA.- mRNA electroporation as a Tool for Immunomonitoring.- mRNA PCR-Based Epitope Chase Method.
