Gene Editing

  1. Variations and Innovations in CRISPR Technology
    1. Cas9 Engineering and Variants
      1. High-Fidelity Cas9 Variants
        1. SpCas9-HF1
          1. eSpCas9
            1. SpRY-Cas9
              1. Reduced Off-target Activity
              2. Cas9 Nickases
                1. Single-Strand Cleavage
                  1. Paired Nickase Strategy
                    1. Reduced Off-target Risk
                    2. Catalytically Dead Cas9
                      1. dCas9 Applications
                        1. DNA Binding without Cleavage
                        2. Miniaturized Cas9 Variants
                          1. Packaging Constraints
                            1. Viral Delivery Applications
                          2. Alternative Cas Proteins
                            1. Cas12 Family
                              1. Cas12a Structure and Function
                                1. Unique Features
                                  1. T-rich PAM Requirements
                                    1. Staggered Cut Generation
                                      1. Trans-cleavage Activity
                                      2. Applications
                                        1. Diagnostics
                                          1. Therapeutic Editing
                                        2. Cas13 Family
                                          1. RNA Targeting Capability
                                            1. Cas13a Mechanism
                                              1. Cas13d Applications
                                                1. RNA Editing and Regulation
                                                  1. Diagnostic Applications
                                                    1. SHERLOCK Platform
                                                  2. Cas14 and Other Compact Systems
                                                    1. Size Advantages
                                                      1. Delivery Benefits
                                                    2. Base Editing Technologies
                                                      1. Cytosine Base Editors
                                                        1. BE3 and BE4max Systems
                                                          1. Cytidine Deaminase Fusion
                                                            1. C-to-T Conversion Mechanism
                                                              1. Applications in Disease Correction
                                                              2. Adenine Base Editors
                                                                1. ABE7.10 and ABE8e Systems
                                                                  1. Adenosine Deaminase Engineering
                                                                    1. A-to-G Conversion Mechanism
                                                                      1. Expanded Targeting Range
                                                                      2. Base Editing Advantages
                                                                        1. No Double-Strand Breaks
                                                                          1. Higher Efficiency than HDR
                                                                            1. Reduced Indel Formation
                                                                            2. Base Editing Limitations
                                                                              1. Limited Editing Window
                                                                                1. Bystander Editing
                                                                                  1. Off-target RNA Editing
                                                                                2. Prime Editing
                                                                                  1. Prime Editing Mechanism
                                                                                    1. Nickase Cas9 Fusion
                                                                                      1. Reverse Transcriptase Integration
                                                                                        1. Prime Editing Guide RNA Design
                                                                                        2. Prime Editing Capabilities
                                                                                          1. Insertions up to 80 bp
                                                                                            1. Deletions
                                                                                              1. All Base Conversions
                                                                                                1. Indel Correction
                                                                                                2. Comparison with Other Methods
                                                                                                  1. Precision vs. HDR
                                                                                                    1. Efficiency vs. Base Editing
                                                                                                      1. Versatility Advantages
                                                                                                      2. Prime Editing Optimization
                                                                                                        1. pegRNA Design
                                                                                                          1. Nicking Strategy
                                                                                                            1. Delivery Considerations
                                                                                                          2. Epigenome Editing
                                                                                                            1. CRISPR Interference
                                                                                                              1. dCas9-KRAB Fusions
                                                                                                                1. Transcriptional Repression
                                                                                                                  1. Chromatin Modifications
                                                                                                                  2. CRISPR Activation
                                                                                                                    1. dCas9-VP64 Systems
                                                                                                                      1. SunTag and SAM Systems
                                                                                                                        1. Transcriptional Enhancement
                                                                                                                        2. Epigenetic Modifications
                                                                                                                          1. DNA Methylation Editing
                                                                                                                            1. Histone Modification Editing
                                                                                                                              1. Chromatin Remodeling
                                                                                                                              2. Applications in Research
                                                                                                                                1. Gene Function Studies
                                                                                                                                  1. Disease Modeling
                                                                                                                                    1. Therapeutic Development

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                                                                                                                                5. The CRISPR-Cas9 System in Detail

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                                                                                                                                7. Gene Editing Workflow and Implementation