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This volume details cutting-edge methods that enables engineering of novel functions in mammalian cells. Chapters guide readers through cell-to-cell communication, cell fate control, protein and RNA-based biosensors together with tools for more reliable and faster mammalian genome editing. 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 key tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Mammalian Synthetic Systems aims to ensure successful results in the further study of this vital field.
List of contents
A directed evolution protocol for engineering minimal transcription factors, based on CIS display.- Set-up and applications of Modular Protein Expression Toolboxes (MoPET) for mammalian systems.- Coiled-coiled interaction toolbox for engineering mammalian cells.- A mammalian-based synthetic biology toolbox to engineer membrane-membrane interfaces.- Engineered protease-responsive RNA Binding Proteins (RBPs) to expand the toolbox of synthetic circuits in mammalian cells.- Mechanistic model-driven biodesign in mammalian synthetic biology.- Realizing Antithetic Integral Feedback Control in Mammalian Cells.- A computational modeling approach for the design of genetic control systems that respond to transcriptional activity.- Flux Balance Analysis of Mammalian Cell Systems.- Multiplex functional characterization of protein variant libraries in mammalian cells with single-copy genomic integration and high throughput DNA sequencing.- Flow Cytometry Quantification of Transient Transfections in Mammalian CellsRNA switches using Cas proteins.- Multiplexed transactivation of mammalian cells using dFnCas12a-VPR.- Anti-CRISPR Proteins and their Application to Control CRISPR-effectors in Mammalian Systems.- Post-translational Remote Control Mediated by Physical Inducers for Rapid Protein Release in Engineered Mammalian Cells.- Detection of microRNAs Using Synthetic Toehold Switch in Mammalian Cells.- Detection of microRNAs Using Synthetic Toehold Switch in Mammalian Cells.- High-throughput spectroscopic analysis of mRNA capping level.- In vitro generation of megakaryocytes from engineered mouse embryonic stem cells.- Preparation of chromobodies for the detection of cell surface epitopes.
