An optimized optogenetic clustering tool for probing protein interaction and function

Taslimi, Amir; Vrana, Justin D.; Chen, Daniel; Borinskaya, Sofya; Mayer, Bruce J.; Kennedy, Matthew J.; Tucker, Chandra L.

doi:10.1038/ncomms5925

Cited by 316 publications

(394 citation statements)

References 44 publications

(60 reference statements)

Supporting

Mentioning

356

Contrasting

Unclassified

Order By: Relevance

“…The slow photoswitches could be used to avoid some unwanted side effects caused by continuous irradiation, such as phototoxicity and unfocused activation owing to sample drift. Recently, methods to improve switch-off kinetics of blue light photoreceptors have been developed using bacterial or yeast colonies-based high throughput screening systems 28,29 . These screening methods can help to further accelerate or decelerate the switch-off kinetics of the present photoswitches.…”

Section: Discussionmentioning

confidence: 99%

Engineered pairs of distinct photoswitches for optogenetic control of cellular proteins

et al. 2015

View full text Add to dashboard Cite

Optogenetic methods take advantage of photoswitches to control the activity of cellular proteins. Here, we completed a multi-directional engineering of the fungal photoreceptor Vivid to develop pairs of distinct photoswitches named Magnets. These new photoswitches were engineered to recognize each other based on the electrostatic interactions, thus preventing homodimerization and enhancing light-induced heterodimerization. Furthermore, we tuned the switch-off kinetics by four orders of magnitude and developed several variants, including those with substantially faster kinetics than any of the other conventional dimerization-based blue spectrum photoswitches. We demonstrate the utility of Magnets as powerful tools that can optogenetically manipulate molecular processes in biological systems.

show abstract

Section: Discussionmentioning

confidence: 99%

Engineered pairs of distinct photoswitches for optogenetic control of cellular proteins

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Interestingly, CRY2-CIBN binding outcompetes CRY2 oligomerization under the same light-activating conditions (Che et al, 2015). Both CRY2-CIBN heterodimerization (Boulina et al, 2013;Hughes et al, 2012;Idevall-Hagren et al, 2012;Kakumoto and Nakata, 2013;Kennedy et al, 2010;Konermann et al, 2013;Lee et al, 2014;Liu et al, 2012; and CRY2 homo-oligomerization (Bugaj et al, 2013;Chang et al, 2014;Taslimi et al, 2014;Wend et al, 2014) have been used for optogenetic control of signal transduction. It appears that CRY2-CIBN-induced protein dimerization mimics the native interaction between the two proteins better .…”

Section: Introductionmentioning

confidence: 99%

Reversible optogenetic control of kinase activity during differentiation and embryonic development

et al. 2016

View full text Add to dashboard Cite

A limited number of signaling pathways are repeatedly used to regulate a wide variety of processes during development and differentiation. The lack of tools to manipulate signaling pathways dynamically in space and time has been a major technical challenge for biologists. Optogenetic techniques, which utilize light to control protein functions in a reversible fashion, hold promise for modulating intracellular signaling networks with high spatial and temporal resolution. Applications of optogenetics in multicellular organisms, however, have not been widely reported. Here, we create an optimized bicistronic optogenetic system using Arabidopsis thaliana cryptochrome 2 (CRY2) protein and the N-terminal domain of cryptochrome-interacting basic-helix-loop-helix (CIBN). In a proofof-principle study, we develop an optogenetic Raf kinase that allows reversible light-controlled activation of the Raf/MEK/ERK signaling cascade. In PC12 cells, this system significantly improves lightinduced cell differentiation compared with co-transfection. When applied to Xenopus embryos, this system enables blue lightdependent reversible Raf activation at any desired developmental stage in specific cell lineages. Our system offers a powerful optogenetic tool suitable for manipulation of signaling pathways with high spatial and temporal resolution in a wide range of experimental settings.

show abstract

“…Recent protein purification studies along with multiangle light scattering (SEC-MALS) experiments have shown that Cry2PHR most likely forms a tetramer (63), which may be subject to higher-order oligomerization. Recently, an E490G mutation in the protein has been shown to greatly enhance the amount of clustering in the Cry2 system (64). It was shown that 40–90% of this cytosolic protein, named Cry2olig, immediately formed clusters when exposed to blue light.…”

Section: Optogenetic Systemsmentioning

confidence: 99%

“…The Cry2/CIBN and Cry2 modules have also found application in several other cellular biological phenomena, including, but not limited to, signal transduction, transcription, and protein-protein interactions (39, 62, 64, 65). …”

Section: Optogenetic Systemsmentioning

confidence: 99%

At Light Speed: Advances in Optogenetic Systems for Regulating Cell Signaling and Behavior

Repina

Rosenbloom

Mukherjee

et al. 2017

Annu. Rev. Chem. Biomol. Eng.

135

125

View full text Add to dashboard Cite

Cells are bombarded by extrinsic signals that dynamically change in time and space. Such dynamic variations can exert profound effects on behaviors, including cellular signaling, organismal development, stem cell differentiation, normal tissue function, and disease processes such as cancer. Although classical genetic tools are well suited to introduce binary perturbations, new approaches were necessary to investigate how dynamic signal variation may regulate cell behavior. This fundamental question is increasingly being addressed with optogenetics, a field focused on engineering and harnessing light-sensitive proteins to interface with cellular signaling pathways. Channelrhodopsins defined optogenetics; however, through recent use of light-responsive proteins with myriad spectral and functional properties, practical applications of optogenetics currently encompass cell signaling, subcellular localization, and gene regulation. Now, important questions regarding signal integration within branch points of signaling networks, asymmetric cell responses to spatially restricted signals, and effects of signal dosage versus duration can be addressed. This review summarizes emerging technologies and applications within the expanding field of optogenetics.

show abstract

An optimized optogenetic clustering tool for probing protein interaction and function

Cited by 316 publications

References 44 publications

Engineered pairs of distinct photoswitches for optogenetic control of cellular proteins

Engineered pairs of distinct photoswitches for optogenetic control of cellular proteins

Reversible optogenetic control of kinase activity during differentiation and embryonic development

At Light Speed: Advances in Optogenetic Systems for Regulating Cell Signaling and Behavior

Contact Info

Product

Resources

About