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This detailed second edition volume expands upon the prior edition by addressing newly emerged technologies as well as improved existing protocols in nucleic acid delivery via nanotechnology. The book addresses topics ranging from chemical synthesis of macromolecules and bioconjugates, novel and established nanoformulations, characterization of these nanoformulations for biophysical, biological and toxicological aspects, and also protocols dealing with application and imaging of such carrier systems in vivo. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Nanotechnology for Nucleic Acid Delivery: Methods and Protocols, Second Edition serves as an ideal guide to researchers seeking to use this vital area of study in their own lab work or to progress knowledge within the field itself.
List of contents
Sequence-Defined Cationic Lipo-Oligomers Containing Unsaturated Fatty Acids for Transfection.- Synthesis of Bioreducible Polycations with Controlled Topologies.- Histidine-Rich Cationic Cell Penetrating Peptides for Plasmid DNA and siRNA Delivery.- Covalent Fluorophore Labeling of Oligonucleotides and Generation of Other Oligonucleotide Bioconjugates.- Synthetic Oligonucleotides in SPECT/CT In Vivo Imaging: Chemical Modifications, In111 Complex Formation, Incorporation into Drug Delivery Systems.- Synthesis of Polyethylenimine-Based Nanocarriers for Systemic Tumor Targeting of Nucleic Acids.- Cationic Photopolymerized Polydiacetylenic (PDA) Micelles for siRNA Delivery.- Lipids for Nucleic Acid Delivery: Cationic or Neutral Lipoplexes, Synthesis, and Particle Formation.- Preparation, Characterization, and In Vitro Evaluation of Lipidoid-Polymer Hybrid Nanoparticles for siRNA Delivery to the Cytosol.- Layer-By-Layer Assembled Nanoparticles for siRNA Delivery.- Layer-By-Layer Film Engineering for Sequential Gene Delivery.- Surface- and Hydrogel-Mediated Delivery of Nucleic Acid Nanoparticles.- In Situ AFM Analysis Investigating Disassembly of DNA Nanoparticles and Nano-Films.- Lyophilization of Synthetic Gene Carriers.- Firefly Luciferase-Based Reporter Gene Assay for Investigating Nanoparticle-Mediated Nucleic Acid Delivery.- Enhancing Nucleic Acid Delivery with Ultrasound and Microbubbles.- Magnetic and Acoustically Active Microbubbles Loaded with Nucleic Acids for Gene Delivery.- Lactate Dehydrogenase Assay for Assessment of Polycation Cytotoxicity.- Combined Fluorimetric Caspase 3/7 Assay and Bradford Protein Determination for Assessment of Polycation-Mediated Cytotoxicity.- Determination of Polycation-Mediated Perturbation of Mitochondrial Respiration in Intact Cells by High-Resolution Respirometry (Oxygraph-2k, OROBOROS).- Evaluating the Regulation of Cytokine Levels after siRNA Treatment in Antigen-Specific Target Cell Populations via Intracellular Staining.- Anti-PEG IgM Production via a PEGylated Nano-Carrier System for Nucleic Acid Delivery.- Near-Infrared Optical Imaging of Nucleic Acid Nanocarriers In Vivo.- Flow Cytometry-Based Cell Type-Specific Assessment of Target Regulation by Pulmonary siRNA Delivery.- Microbubbles for Nucleic Acid Delivery in Liver Using Mild Sonoporation.- Lipopeptide Delivery of siRNA to the Central Nervous System.
Summary
This detailed second edition volume expands upon the prior edition by addressing newly emerged technologies as well as improved existing protocols in nucleic acid delivery via nanotechnology. The book addresses topics ranging from chemical synthesis of macromolecules and bioconjugates, novel and established nanoformulations, characterization of these nanoformulations for biophysical, biological and toxicological aspects, and also protocols dealing with application and imaging of such carrier systems in vivo. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Nanotechnology for Nucleic Acid Delivery: Methods and Protocols, Second Edition serves as an ideal guide to researchers seeking to use this vital area of study in their own lab work or to progress knowledge within the field itself.
