Optogenetic approaches to cell migration and beyond

Weitzman, Matthew D.; Hahn, Klaus M.

doi:10.1016/j.ceb.2014.08.004

Cited by 84 publications

(61 citation statements)

References 60 publications

(75 reference statements)

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“…In addition to small molecule activation of protein function, tools for optical activation of protein function have received significant interest in recent years. 17,18 Small molecule control and light control complement each other and can be synergistically employed in a powerful fashion. 19,20 …”

Section: Introductionmentioning

confidence: 99%

Small-molecule control of protein function through Staudinger reduction

et al. 2016

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Using small molecules to control the function of proteins in live cells with complete specificity is highly desirable, but challenging. Here we report a small molecule switch that can be used to control protein activity. The approach uses a phosphine-mediated Staudinger reduction to activate protein function. Genetic encoding of an ortho-azidobenzyloxycarbonyl amino acid using a pyrrolysyl tRNA synthetase/tRNACUA pair in mammalian cells enables the site-specific introduction of a small molecule-removable protecting group into the protein of interest. Strategic placement of this group renders the protein inactive until deprotection through a bioorthogonal Staudinger reduction delivers the active, wild-type protein. This developed methodology was applied to the conditional control of several cellular processes, including bioluminescence (luciferase), fluorescence (EGFP), protein translocation (nuclear localization sequence), DNA recombination (Cre), and gene editing (Cas9).

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Section: Introductionmentioning

confidence: 99%

Small-molecule control of protein function through Staudinger reduction

et al. 2016

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“…[39] Light-gated ion channels, notably channelrhodopsins, open to allow the flow of ions in response to illumination. Non-channel classes of light actuators and their application to cell motility have been recently reviewed[40]. …”

Section: Controlled Activation Of Protein Activitymentioning

confidence: 99%

Synthetic mechanobiology: engineering cellular force generation and signaling

Hughes

Kumar

2016

Current Opinion in Biotechnology

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“…One such protein is phytochrome B (PhyB) which undergoes a conformational change upon exposure to light of visible red wavelengths (∼650–670 nm) to heterodimerize with the transcription factor phytochrome-interacting factor 3 (PIF3). Unlike other photosensitive proteins, this dimerization can be reverted through exposure to longer wavelengths of light (∼700–750 nm), which induces monoisomerization of PhyB and releases PIF3, allowing for very precise control of protein activity [32,36]. …”

Section: Complex Formationmentioning

confidence: 99%

Targeting protein function: the expanding toolkit for conditional disruption

Campbell

Bennett

2016

Biochemical Journal

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A major objective in biological research is to understand spatial and temporal requirements for any given gene, especially in dynamic processes acting over short periods, such as catalytically driven reactions, subcellular transport, cell division, cell rearrangement and cell migration. The interrogation of such processes requires the use of rapid and flexible methods of interfering with gene function. However, many of the most widely used interventional approaches, such as RNAi or CRISPR (clustered regularly interspaced short palindromic repeats)-Cas9 (CRISPR-associated 9), operate at the level of the gene or its transcripts, meaning that the effects of gene perturbation are exhibited over longer time frames than the process under investigation. There has been much activity over the last few years to address this fundamental problem. In the present review, we describe recent advances in disruption technologies acting at the level of the expressed protein, involving inducible methods of protein cleavage, (in)activation, protein sequestration or degradation. Drawing on examples from model organisms we illustrate the utility of fast-acting techniques and discuss how different components of the molecular toolkit can be employed to dissect previously intractable biochemical processes and cellular behaviours.

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Optogenetic approaches to cell migration and beyond

Cited by 84 publications

References 60 publications

Small-molecule control of protein function through Staudinger reduction

Small-molecule control of protein function through Staudinger reduction

Synthetic mechanobiology: engineering cellular force generation and signaling

Targeting protein function: the expanding toolkit for conditional disruption

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