Gene Editing

  1. Major Gene Editing Technologies
    1. First-Generation Tools
      1. Meganucleases
        1. Structure and DNA Recognition
          1. Protein Fold
            1. DNA Binding Specificity
            2. Mechanism of Action
              1. Target Site Recognition
                1. DNA Cleavage
                2. Applications and Limitations
                  1. Limited Targeting Options
                    1. Engineering Challenges
                  2. Homologous Recombination
                    1. Natural Recombination Mechanisms
                      1. Gene Targeting in Model Organisms
                        1. Efficiency Limitations
                      2. Second-Generation Tools
                        1. Zinc-Finger Nucleases
                          1. Structure and Design
                            1. Zinc-Finger DNA Binding Domains
                              1. Zinc-Finger Motifs
                                1. DNA Recognition Code
                                  1. Modular Assembly
                                  2. FokI Nuclease Domain
                                    1. Dimerization Requirement
                                      1. Cleavage Mechanism
                                    2. Engineering and Optimization
                                      1. Zinc-Finger Assembly Methods
                                        1. Specificity Enhancement
                                        2. Mechanism of Action
                                          1. DNA Targeting Process
                                            1. Double-Strand Break Formation
                                            2. Applications
                                              1. Therapeutic Applications
                                                1. Research Applications
                                                2. Limitations
                                                  1. Design Complexity
                                                    1. Context-dependent Effects
                                                      1. Off-target Concerns
                                                    2. Transcription Activator-Like Effector Nucleases
                                                      1. TALE Protein Origins
                                                        1. Bacterial Plant Pathogens
                                                          1. Natural DNA Binding Function
                                                          2. Structure and Design
                                                            1. TALE DNA Binding Domains
                                                              1. Repeat Variable Diresidues
                                                                1. DNA Recognition Code
                                                                  1. Modular Design Principles
                                                                  2. FokI Nuclease Domain Fusion
                                                                  3. Engineering TALENs
                                                                    1. Assembly Methods
                                                                      1. Optimization Strategies
                                                                      2. Mechanism of Action
                                                                        1. Target Site Recognition
                                                                          1. DNA Binding and Cleavage
                                                                          2. Advantages over ZFNs
                                                                            1. Simplified Design Rules
                                                                              1. Improved Specificity
                                                                                1. Reduced Context Effects
                                                                                2. Applications and Limitations
                                                                                  1. Research Applications
                                                                                    1. Therapeutic Development
                                                                                      1. Remaining Challenges

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                                                                                2. Introduction to Gene Editing

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                                                                                4. The CRISPR-Cas System