Optogenetics Methods and Protocols

This volume explores the latest advancements in the field of optogenetics and how it uses cellular light-sensing components and genetic engineering to control proteins and biological processes. The book chapters are organized into four parts. Part One focuses on intracellular optogenetic components for control of specific cell functions; Part Two looks at externally supplied light regulators that do not require genetic manipulation of target cells; Part Three highlights optogenetic control of organelles, and Part Four introduces technical tools required for light induction in optogenetic experiments, as well as a method for performing and analyzing optogenetic cell-cell adhesion. 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.

Cutting-edge and practical, Optogenetics: Methods and Protocols is a valuable resource to help researchers understand and apply the concepts of optogenetics and the underlying bioengineering principles, and establish the required technical light-illumination setups for administering light inputs and analysis of experimental outcomes.  

Optogenetic Control of B. subtilis Gene Expression using the CcaSR System.- The Yeast Optogenetic Toolkit (yOTK) for Spatiotemporal Control of gene Expression in Budding Yeast.- Optoribogenetic Modulation of Transcription.- A Red Light-Activatable Endogenous Gene Transcription System with Red-CPTS.- Targeted Activation of Programmed Cell Death Pathways by Optogenetics.- Modulating Optogenetic YAP In Vitro and In Vivo.- Design and Use of AsLOV2-Based Optogenetic Tools for Actin Imaging.- Optogenetic Engineered Living Materials.- Light-Controlled Secretion and Injection of Proteins into Eukaryotic Cells by Optogenetic Control of the Bacterial Type III Secretion System.- Synthesis and Application of Photcaged Isopropyl ß-d-1-thiogalactopyranoside for Light-Mediated Control of Bacterial Gene Expression.- Optogenetic Clustering and Stimulation of the T Cell Receptor in Non-Genetically Modified Human T Cells.- Extracellular Optogenetics to Interrogate Unmodified Primary Cell Functions.- Alternating Cellular Functions by Optogenetic Control of Organelles.- Light-Inducible Deformation of Mitochondria in Live Cells.- Chlorophyll-Based Optogenetics to Control Membraneless Organelles.- Automated Plate Reader-Based Assays of Light-Activated GPCRs.- Modular Light-Emitting Diode Shelving Systems for Scalable Optogenetics.- Analysis of Light-Controlled Artificial Cell-Cell Adhesions.