Curated Optogenetic Publication Database

1.

Optogenetics Methods and Protocols

blue green red AsLOV2 CcaS/CcaR Cph1 CRY2/CIB1 CRY2olig DrBphP iLID LOVTRAP Magnets PAL PhyB/PIF6 TtCBD TULIP VVD YtvA 3T3-L1 B. subtilis Cos-7 E. coli H9c2 HaCaT HEK293T HeLa HFF-1 Jurkat MDA-MB-231 MKN28 mouse in vivo primary mouse T cells S. cerevisiae Schneider 2 U-2 OS Y. enterocolitica zebrafish in vivo

Methods Mol Biol, 26 Dec 2024 DOI: 10.1007/978-1-0716-4047-0 Link to full text

Abstract: 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.

2.

Optogenetic activators of apoptosis, necroptosis, and pyroptosis.

blue CRY2olig Caco-2 HaCaT HEK293T HeLa HT-29 MCF7 RAW264.7 U-937 zebrafish in vivo Cell death

J Cell Biol, 14 Apr 2022 DOI: 10.1083/jcb.202109038 Link to full text

Abstract: Targeted and specific induction of cell death in an individual or groups of cells hold the potential for new insights into the response of tissues or organisms to different forms of death. Here, we report the development of optogenetically controlled cell death effectors (optoCDEs), a novel class of optogenetic tools that enables light-mediated induction of three types of programmed cell death (PCD)—apoptosis, pyroptosis, and necroptosis—using Arabidopsis thaliana photosensitive protein Cryptochrome-2. OptoCDEs enable a rapid and highly specific induction of PCD in human, mouse, and zebrafish cells and are suitable for a wide range of applications, such as sub-lethal cell death induction or precise elimination of single cells or cell populations in vitro and in vivo. As the proof-of-concept, we utilize optoCDEs to assess the differences in neighboring cell responses to apoptotic or necrotic PCD, revealing a new role for shingosine-1-phosphate signaling in regulating the efferocytosis of the apoptotic cell by epithelia.

3.

An optogenetic method for interrogating YAP1 and TAZ nuclear-cytoplasmic shuttling.

blue LOVTRAP HaCaT Signaling cascade control

J Cell Sci, 1 Jun 2021 DOI: 10.1242/jcs.253484 Link to full text

Abstract: The shuttling of transcription factors and transcriptional regulators into and out of the nucleus is central to the regulation of many biological processes. Here we describe a new method for studying the rates of nuclear entry and exit of transcriptional regulators. A photo-responsive AsLOV (Avena sativa Light Oxygen Voltage) domain is used to sequester fluorescently-labelled transcriptional regulators YAP1 and TAZ/WWTR1 on the surface of mitochondria and reversibly release them upon blue light illumination. After dissociation, fluorescent signals from mitochondria, cytoplasm and nucleus are extracted with a bespoke app and used to generate rates of nuclear entry and exit. Using this method, we demonstrate that phosphorylation of YAP1 on canonical sites enhances its rate of nuclear export. Moreover, we provide evidence that, despite high intercellular variability, YAP1 import and export rates correlated within the same cell. By simultaneously releasing YAP1 and TAZ from sequestration, we show that their rates of entry and exit are correlated. Furthermore, combining the optogenetic release of YAP1 with lattice light-sheet microscopy revealed high heterogeneity of YAP1 dynamics within different cytoplasmic regions, demonstrating the utility and versatility of our tool to study protein dynamics.