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 control of YAP cellular localisation and function.

blue AsLOV2 HEK293T HFF-1 MKN28 zebrafish in vivo Signaling cascade control

EMBO Rep, 25 Jul 2022 DOI: 10.15252/embr.202154401 Link to full text

Abstract: YAP, an effector of the Hippo signalling pathway, promotes organ growth and regeneration. Prolonged YAP activation results in uncontrolled proliferation and cancer. Therefore, exogenous regulation of YAP activity has potential translational applications. We present a versatile optogenetic construct (optoYAP) for manipulating YAP localisation, and consequently its activity and function. We attach a LOV2 domain that photocages a nuclear localisation signal (NLS) to the N-terminus of YAP. In 488 nm light, the LOV2 domain unfolds, exposing the NLS, which shuttles optoYAP into the nucleus. Nuclear import of optoYAP is reversible and tuneable by light intensity. In cell culture, activated optoYAP promotes YAP target gene expression and cell proliferation. Similarly, optofYap can be used in zebrafish embryos to modulate target genes. We demonstrate that optoYAP can override a cell's response to substrate stiffness to generate anchorage-independent growth. OptoYAP is functional in both cell culture and in vivo, providing a powerful tool to address basic research questions and therapeutic applications in regeneration and disease.

3.

Optogenetic control of cell morphogenesis on protein micropatterns.

blue AsLOV2 HFF-1 Control of cytoskeleton / cell motility / cell shape

bioRxiv, 4 Mar 2019 DOI: 10.1101/563353 Link to full text

Abstract: Cell morphogenesis is critical for embryonic development, tissue formation, and wound healing. Our ability to manipulate endogenous mechanisms to control cell shape, however, remains limited. Here we combined surface micropatterning of adhesion molecules with optogenetic activation of intracellular signaling pathways to control the nature and morphology of cellular protrusions. We employed geometry-dependent pre-organization of cytoskeletal structures together with acute activation of signaling pathways that control actin assembly to create a tool capable of generating membrane protrusions at defined cellular locations. Further, we find that the size of microfabricated patterns of adhesion molecules influences the molecular mechanism of cell protrusion: larger patterns enable cells to create actin-filled lamellipodia while smaller patterns promote formation of spherical blebs. Optogenetic perturbation of signaling pathways in these cells changes the size of blebs and convert them into lamellipodia. Our results demonstrate how the coordinated manipulation of adhesion geometry and cytoskeletal dynamics can be used to control membrane protrusion and cell morphogenesis.