Metabolic Engineering

  1. Tools for Genetic Modification
    1. Recombinant DNA Technology
      1. Plasmids and Vectors
        1. High-Copy Vectors
          1. Low-Copy Vectors
            1. Shuttle Vectors
              1. Vector Design Considerations
              2. Restriction Enzymes and Ligation
                1. Type II Restriction Enzymes
                  1. Blunt-End Ligation
                    1. Sticky-End Ligation
                      1. Golden Gate Assembly
                      2. Polymerase Chain Reaction
                        1. PCR Principles
                          1. Primer Design
                            1. Applications in Cloning
                              1. Quantitative PCR
                            2. Gene Expression Control
                              1. Promoters
                                1. Constitutive Promoters
                                  1. Inducible Promoters
                                    1. Repressible Promoters
                                      1. Promoter Strength Tuning
                                        1. Promoter Libraries
                                        2. Ribosome Binding Sites
                                          1. RBS Sequence Design
                                            1. Translation Initiation Rate Control
                                              1. RBS Calculator Tools
                                              2. Terminators
                                                1. Transcription Termination Mechanisms
                                                  1. Intrinsic Terminators
                                                    1. Rho-Dependent Terminators
                                                      1. Terminator Strength
                                                    2. Gene Knockout and Knockdown
                                                      1. Homologous Recombination
                                                        1. Double Crossover Events
                                                          1. Single Crossover Events
                                                            1. Markerless Deletion Strategies
                                                              1. Recombineering
                                                              2. RNA Interference
                                                                1. Mechanism of RNAi
                                                                  1. siRNA Design
                                                                    1. Applications in Eukaryotes
                                                                    2. Antisense RNA
                                                                      1. Mechanism of Action
                                                                        1. Design Considerations
                                                                      2. Genome Editing Technologies
                                                                        1. CRISPR-Cas Systems
                                                                          1. Mechanism of CRISPR-Cas9
                                                                            1. Guide RNA Design
                                                                              1. PAM Sequence Requirements
                                                                                1. Off-Target Effects
                                                                                  1. Base Editing
                                                                                    1. Prime Editing
                                                                                    2. Zinc-Finger Nucleases
                                                                                      1. Structure and DNA Recognition
                                                                                        1. Zinc-Finger Design
                                                                                          1. Applications and Limitations
                                                                                          2. TALENs
                                                                                            1. DNA Binding Specificity
                                                                                              1. TALE Repeat Engineering
                                                                                                1. Comparison to Other Technologies
                                                                                              2. Directed Evolution and Mutagenesis
                                                                                                1. Error-Prone PCR
                                                                                                  1. Principles and Protocols
                                                                                                    1. Mutation Rate Control
                                                                                                      1. Library Generation
                                                                                                      2. DNA Shuffling
                                                                                                        1. Recombination of Homologous Genes
                                                                                                          1. Family Shuffling
                                                                                                            1. Diversity Generation
                                                                                                            2. Site-Directed Mutagenesis
                                                                                                              1. Primer Design
                                                                                                                1. Targeted Amino Acid Changes
                                                                                                                  1. Saturation Mutagenesis
                                                                                                                  2. Chemical Mutagenesis
                                                                                                                    1. Mutagen Types
                                                                                                                      1. Mutagenesis Protocols

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                                                                                                                  2. Fundamentals of Cellular Metabolism

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                                                                                                                  4. Analytical Methods for Metabolic Engineering