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spatiotemporal control of Protein Labeling in Live Cells Enabled by a Photoreactive Ligand-Directed Probe

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Spatiotemporal Control of Protein Labeling in Live Cells Ena...

Intracavity spatiotemporal metasurfaces

第十一届全国化学生物学学术会议

作者： Huaibin Yu Yu Fu Shuangshuang Wang Yuanjie Sun Fangrui Zhong Yuzhou Wu School of Chemistry and Chemical Engineering Huazhong University of Science and Technology （HUST）

Selective protein labeling in live cells has been crucial for molecular biology studies[1-3]. However, due to the highly complicated environment in live cells with coexistence of thousands of proteins, it represents a formidable challenge to fish out a specific protein of interest（POI） from this vast pool. Particularly, there are many scenarios that the POI might need to be labeled not only in a site-specific and target-selective fashion, but also with a handle to control the labeling time point in order to study the dynamic function and location of a protein[4-5]. Herein we develop a spatiotemporal controlled probe that enables the site-selective reaction of target proteins both in vitro and live cell conditions, upon time controlled photodecaging. The probe was consisted of a moderately reactive 3-（hydroxymethyl）-2-naphthol derivatives（oNQMs） assisted by ligand-binding affinity chemistry, allowed us to selectively modify natural proteins proximal to the ligand-binding pocket once light triggered in mild conditions. Besides, the photolabile（2-nitrophenyl） propyloxycarbonyl（NPPOC） was also employed to acts as the leaving group, which could be cleaved under photolytic conditions. Using this new approach, we detected that the probe can realize noninvasive spatiotemporal controlled selective labeling on the live MCF-7 cell surface and cytoplasm, whereas the genetic manipulations have been typically of necessity with previous methods.

关键词： Selective protein labeling protein of interest spatiotemporal control photodecaging

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Advanced Photonics 2023年第2期5卷 46-53页

作者： Wenhe Jia Chenxin Gao Yongmin Zhao Liu Li Shun Wen Shuai Wang Chengying Bao Chunping Jiang Changxi Yang Yuanmu Yang Tsinghua University State Key Laboratory of Precision Measurement Technology and InstrumentsDepartment of Precision InstrumentBeijingChina Chinese Academy of Sciences Suzhou Institute of Nano-Tech and Nano-BionicsKey Laboratory of Nanodevices and ApplicationsSuzhouChina

Optical metasurfaces are endowed with unparallel flexibility to manipulate the light field with a subwavelength spatial *** metasurfaces to materials with strong optical nonlinearity may allow ultrafast spatiotemporal light field ***,most metasurfaces demonstrated thus far are linear ***,we experimentally demonstrate simultaneous spatiotemporal laser mode control using a single-layer plasmonic metasurface strongly coupled to an epsilon-near-zero(ENZ)material within a fiber laser *** the geometric phase of the metasurface is utilized to convert the laser’s transverse mode from a Gaussian beam to a vortex beam carrying orbital angular momentum,the giant nonlinear saturable absorption of the ENZ material enables pulsed laser generation via the Q-switching *** direct integration of a spatiotemporal metasurface in a laser cavity may pave the way for the development of miniaturized laser sources with tailored spatial and temporal profiles,which can be useful for numerous applications,such as superresolution imaging,high-density optical storage,and three-dimensional laser lithography.

关键词： spatiotemporal control metasurface epsilon-near-zero strong coupling.

Design and mathematical analysis of activating transcriptional amplifiers that enable modular temporal control in synthetic juxtacrine circuits

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Synthetic and Systems Biotechnology 2023年第4期8卷 654-672页

作者： Calvin Lam Department of Biochemistry and Molecular Biology University of Nebraska Medical CenterOmahaNE68198USA.

The ability to control mammalian cells such that they self-organize or enact therapeutic effects as desired has incredible *** only would it further our understanding of native processes such as development and the immune response,but it would also have powerful applications in medical fields such as regenerative medicine and *** control is typically obtained by synthetic circuits that use synthetic receptors,but control remains *** synthetic juxtacrine receptors(SJRs)are widely used as they are fully modular and enable spatial control,but they have limited gene expression amplification and temporal *** these are integral facets to cell control,I therefore designed transcription factor based amplifiers that amplify gene expression and enable unidirectional temporal control by prolonging duration of target gene *** a validated in silico framework for SJR signaling,I combined these amplifiers with SJRs and show that these SJR amplifier circuits can direct spatiotemporal patterning and improve the quality of self-organization.I then show that these circuits can improve chimeric antigen receptor(CAR)T cell tumor killing against various heterogenous antigen expression *** amplifiers are flexible tools that improve control over SJR based circuits with both basic and therapeutic applications.

关键词： Synthetic biology Synthetic development Synthetic immunotherapy synNotch SNIPR Amplifiers Self-organization Temporal control spatiotemporal control CAR T cell Synthetic receptors

Conditional gene expression in invertebrate animal models

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Journal of Genetics and Genomics 2021年第1期48卷 14-31页

作者： Brecht Driesschaert Lucas Mergan Liesbet Temmerman Animal Physiology and Neurobiology Department of BiologyUniversity of Leuven(KU Leuven)Naamsestraat 59-Box 2465B-3000 LeuvenBelgium

A mechanistic understanding of biology requires appreciating spatiotemporal aspects of gene expression and its functional *** expression allows for (ir)reversible switching of genes on or off,with the potential of spatial and/or temporal *** provides a valuable complement to the more often used constitutive gene (in)activation through mutagenesis,providing tools to answer a wider array of research questions across biological *** and/or temporal control are granted primarily by(combinations of) specific promoters,temperature regimens,compound addition,or *** use of such genetic tool kits is particularly widespread in invertebrate animal models because they can be applied to study biological processes in short time frames and on large scales,using organisms amenable to easy genetic *** years witnessed an exciting expansion and optimization of such tools,of which we provide a comprehensive overview and discussion regarding their use in *** mechanism,applicability,benefits,and drawbacks of each of the systems,as well as further developments to be expected in the foreseeable future,are highlighted.

关键词： Conditional gene expression spatiotemporal control Invertebrate Model organism Drosophila melanogaster Caenorhabditis elegans

Growth factor delivery using extracellular matrix-mimicking substrates for musculoskeletal tissue engineering and repair

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Bioactive Materials 2021年第7期6卷 1945-1956页

作者： Robert C.H.Gresham Chelsea S.Bahney J.Kent Leach UC Davis Department of Biomedical EngineeringDavisCAUSA Steadman Phillippon Research Institute VailCOUSA UCSF Orthopaedic Trauma Institute San FranciscoCAUSA UC Davis Health Department of Orthopaedic SurgeryDavisCAUSA

Therapeutic approaches for musculoskeletal tissue regeneration commonly employ growth factors(GFs)to influence neighboring cells and promote migration,proliferation,or *** promising results in preclinical models,the use of inductive biomacromolecules has achieved limited success in translation to the *** field has yet to sufficiently overcome substantial hurdles such as poor spatiotemporal control and supraphysiological dosages,which commonly result in detrimental side *** presentation and retention of biomacromolecules is regulated by the extracellular matrix(ECM),which acts as a reservoir for GFs via electrostatic *** in the manipulation of extracellular proteins,decellularized tissues,and synthetic ECM-mimetic applications across a range of biomaterials have increased the ability to direct the presentation of *** application of biomaterial technologies utilizing ECM mimetics increases tissue regeneration without the reliance on supraphysiological doses of inductive *** review describes recent strategies to manage GF presentation using ECM-mimetic substrates for the regeneration of bone,cartilage,and muscle.

关键词： Growth factor Extracellular matrix spatiotemporal control Tissue engineering Affinity

Physical stimuli-responsive cell-free protein synthesis

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Synthetic and Systems Biotechnology 2020年第4期5卷 363-368页

作者： Junzhu Yang Yuan Lu Key Laboratory of Industrial Biocatalysis Ministry of EducationDepartment of Chemical EngineeringTsinghua UniversityBeijingChina

Cell-free protein synthesis has been developed as a critical platform in synthetic *** the cell-based synthesis system,cell-free system activates transcriptional and translational mechanisms in vitro,and can control protein synthesis by artificially adding components or ***,the control method puts forward higher requirements in terms of accurate and non-toxic control,which cannot be achieved by chemical *** cell-free system,physical signal is a kind of ideal spatiotemporal control approach to replace chemical substances,realizing high accuracy with little side *** we review the methods of using physical signals to control gene expression in cell-free systems,including studies based on light,temperature,electric field,and magnetic *** transfer of these switches into cell-free system further expands the flexibility and controllability of the system,thus further expanding the application capability of cell-free ***,existing problems such as signal source and signal transmission are discussed,and future applications in pharmaceutical production,delivery and industrial production are further looked into.

关键词： Cell-free system Physical stimuli spatiotemporal control Synthetic biology