Lentiviral Vectors and Exosomes as Gene and Protein Delivery Tools

ISBN-13: 9781493937516 / Angielski / Twarda / 2016 / 276 str.

Maurizio Federico

Opis

This volume provides current methods and protocols for gene and protein delivery based on both lentivirus-generated and spontaneously released nanovesicles. Lentiviral Vectors and Exosomes as Gene and Protein Delivery Tools guides readers through methods on macromolecule delivery and chapters describe the LV-based protocols of gene engineering. Written in the highly 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 laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.Authoritative and cutting-edge, Lentiviral Vectors and Exosomes as Gene and Protein Delivery Tools aims to ensure successful results in the further study of this vital field.

Spis treści

Construction of Modular Lentiviral Vectors for Effective Gene Expression and Knockdown.- Development of Inducible Molecular Switches Based on All-In-One Lentiviral Vectors Equipped With Drug Controlled FLP Recombinase.- Production Of Retrovirus-Based Vectors In Mildly Acidic Ph Conditions.- Optimized Lentiviral Transduction Protocols by use of a Poloxamer Enhancer, Spinoculation and Scfv-Antibody Fusions To VSV-G.- Transduction of Murine Hematopoietic Stem Cells with Tetracycline-Regulated Lentiviral Vectors.- Introduction of Shrnas, Mirnas or Antagomirs into Primary Human Liver Cells Through Lentiviral Vectors.- Production and Concentration of Lentivirus for Transduction of Primary Human T Cells.- Generating Transgenic Mice by Lentiviral Transduction of Spermatozoa Followed by In Vitro Fertilization and Embryo Transfer.- The LAM-PCR Method to Sequence LV Integration Sites.- Conditional Rnai using the Lentiviral GLTR System.- Lentiviral Vectors for the Engineering of Implantable Cells Secreting Recombinant Antibodies.- Transient Expression of Green Fluorescent Protein in Integrase-Defective Lentiviral Vector Transduced 293T Cell Line.- Intrastriatal Delivery of Integration-Deficient Lentiviral Vectors in a Rat Model of Parkinson's Disease.- Development of Lentiviral Vectors for Targeted Integration and Protein Delivery.- Biogenesis and Functions of Exosomes and Extracellular Vesicles.- Generation, Quantification, and Tracing of Metabolically Labeled Fluorescent Exosomes.- Cardiac Myocyte Exosome Isolation.- Incorporation of Heterologous Proteins in Engineered Exosomes.- Exosome-Mediated Targeted Delivery of Mirnas.