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This volume details methods that will aid in the selection of promoter sequences and vector components and methods for the assembly and testing of synthetic promoters with examples of their application. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Mammalian Synthetic Promoters: Methods and Protocols aims to aid researchers that are new to the field of synthetic promoters and inspire new developments.
Initial Considerations before Designing a Promoter Construct.- Demonstrating Interactions of Transcription Factors with DNA by Electrophoretic Mobility Shift Assay.- Chromatin Immunoprecipitation and Quantitative Real-time PCR to Assess Binding of a Protein of Interest to Identified Predicted Binding Sites within a Promoter.- Secreted Reporters for Monitoring Multiple Promoter Function.- Bioluminescence Monitoring Of Promoter Activity In Vitro and In Vivo.- Monitoring Promoter Activity by Flow Cytometry.- Functional Screening of Core Promoter Activity.- Bioinformatically Informed Design of Synthetic Mammalian Promoters.- Synthetic Tumor Specific Promoters for Transcriptional Regulation of Viral Replication.- Constructing Strong Cell-Type-Specific Promoters through Informed Design.- PCR Assembly of Synthetic Promoters.- The Tetracycline Responsive System.- Light-responsive Promoters.- A Simple Method for Constructing Artificial Promoters Activated in Response to Ultrasound Stimulation.- Promoter Activation with Electromagnetism.- Application of Synthetic Tumor Specific Promoters Responsive to the Tumor Microenvironment.- A Modular Approach to Building Complex Synthetic Circuits.- Computational Sequence Design with R2oDNA Designer.- Design of Synthetic Promoters for Gene Circuits in Mammalian Cells.
