Full Description
This detailed collection explores methods that leverage and extend the capabilities of synthetic biology, guiding the reader from foundational molecular manipulations to complex cellular applications. Beginning with chapters on engineering proteins and enzymes, the book continues by covering methods for the repurposing of viral architectures, from bacteriophages to virus-like particles, as programmable nanoscaffolds, CRISPR-driven genome engineering, directed evolution and selection, as well as the synthesis of functional genetic libraries and the groundbreaking technology of biological "memory," where a cell's history is permanently recorded into its DNA. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Biological Macromolecule Engineering Technologies serves as an ideal guide for researchers looking to drive the next wave of discovery in biological macromolecule engineering and address some of humanity's most pressing challenges in health, energy, and manufacturing.
Contents
High Level Protein Expression in Pseudomonas putida Using the Phage-Based pPUT Expression System.- New-to-Nature Photoenzymatic Catalysis.- CRISPR-Hybrid: Intracellular Selection of CRISPR-Associated Aptamers in Bacteria.- Designing and Engineering Enzymes for New-to-Nature Photobiocatalysis.- A Yeast Two-Hybrid Assay to Develop Optogenetic Tools for Controlling Protein-Protein Interactions.- CRISPR-Engineered Bacteriophage T4 for Foot-and-Mouth Disease Nanoparticle Vaccine Development.- Endoskeleton-Enhanced Virus-Like Particles (VLPs): Internal Cross-Linking of Physalis Mottle Virus and Stability Tests.- An Integrated Workflow for Phage Selection, Amplification, and Surface Modification.- CRISPR-Cpf1-Mediated T4 Phage Genome Editing for One-Step In Vivo Display of Heterologous Protein.- Methods for Applying Prime Editing and Inverse Prime Editing in Mammalian Cell Culture.- Constructing Drive-and-Process (DAP) CRISPR Guide RNA Arrays for Multiplexed Base- and Prime-Editing.- Production of Cell Models with Differential Zygosity Using CRISPR Base Editors.- Phage-Assisted Continuous Selection of Bioactive Cyclic Peptides.- Targeted Transition Mutations in Phage-Assisted Evolution.- Phage Display In Vivo: Protocols for Identifying Brain-Targeting Peptides in Mouse Models of Neurodegeneration.- Generation of Diversely Functionalized DNA Libraries Using Ligase-Catalyzed Oligonucleotide Polymerization (LOOPER).- Solid Phase Chemical Synthesis of RNA.- Fluorescence-Based Assays to Evaluate the Functionality of Phage-Derived Regulatory Elements for Applications in Synthetic Biology.- Using peCHYRON to Record Mammalian Cells' History of Chemical Exposure into Their DNA.
