Recent Advances in Fluorescence Imaging by Genetically Encoded Non-canonical Amino Acids

Lee, Sanghee; Kim, Jonghoon; Koh, Minseob

doi:10.1016/j.jmb.2021.167248

Cited by 19 publications

(14 citation statements)

References 113 publications

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“…On the contrary, the sitespecific incorporation of ncAAs into peptides or proteins can be used to create materials with new chemical properties, architecture, and bioactivities that are beyond natural limits. These materials have been used for various applications, such as isotopic [100] and fluorescent labeling, [101,102] bioconjugation, [103] and for the production of novel enzymes and therapeutics. [104][105][106][107][108] It is, therefore, not very surprising that immense efforts have been made to expand the nature's repertoire of ribosomal monomers, particularly structures bearing backbones or moieties significantly distinct from those of canonical amino acids.…”

Section: Production Of Biobased Materials Containing Noncanonical Ami...mentioning

confidence: 99%

Programmable Synthesis of Biobased Materials Using Cell‐Free Systems

et al. 2022

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Motivated by the intricate mechanisms underlying biomolecule syntheses in cells that chemistry is currently unable to mimic, researchers have harnessed biological systems for manufacturing novel materials. Cell‐free systems (CFSs) utilizing the bioactivity of transcriptional and translational machineries in vitro are excellent tools that allow supplementation of exogenous materials for production of innovative materials beyond the capability of natural biological systems. Herein, recent studies that have advanced the ability to expand the scope of biobased materials using CFS are summarized and approaches enabling the production of high‐value materials, prototyping of genetic parts and modules, and biofunctionalization are discussed. By extending the reach of chemical and enzymatic reactions complementary to cellular materials, CFSs provide new opportunities at the interface of materials science and synthetic biology.

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Section: Production Of Biobased Materials Containing Noncanonical Ami...mentioning

confidence: 99%

Programmable Synthesis of Biobased Materials Using Cell‐Free Systems

et al. 2022

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“…Coined bioorthogonal reactions, these ligations often fullfill the criteria of click chemistry as they are high-yielding and exhibit fast kinetics, and mostly belong to the cycloaddition family. Bioorthogonal chemistry has been applied successfully to diverse biological models over the past 20 years − and has greatly facilitated in vivo , ex vivo , and in situ studies of the main classes of biomolecules such as glycans, , lipids, nucleic acids, and proteins exploiting a wide range of bioanalytical and bioimaging readout techniques. , Although many notable examples have been described in mammalian living systems, , there are surprisingly few reports in plants, − despite the fact that they represent 82% of all living matter on earth and make up the first economic resource in terms of nutrition, renewable materials and renewable energies. Lignin is one of the main constituents of plant cell walls, with cellulose.…”

Section: Introductionmentioning

confidence: 99%

Exploring Lignification Complexity in Plant Cell Walls with Airyscan Super-resolution Microscopy and Bioorthogonal Chemistry

Simon

Morel

Neutelings

et al. 2023

Chemical & Biomedical Imaging

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In this paper, we present the use of multiplex click/bioorthogonal chemistry combined with super-resolution Airyscan microscopy to track biomolecules in living systems with a focus on studying lignin formation in plant cell walls. While laser scanning confocal microscopy (LSCM) provided insights into the tissue-scale dynamics of lignin formation and distribution in our previous reports, its limited resolution precluded an in-depth analysis of lignin composition at the unique cell wall or substructure level. To overcome this limitation, we explored the use of Airyscan microscopy, which, among the super-resolution techniques available, offers an optimal balance between performance, cost, accessibility, and ease of implementation. Our study demonstrates that a triple labeling strategy using copper-catalyzed azide–alkyne cycloaddition (CuAAC), strain-promoted azide–alkyne cycloaddition (SPAAC), and inverse electronic-demand Diels–Alder cycloaddition (IEDDA) to label modified lignin metabolic precursors can be combined with Airyscan microscopy to reveal the zones of active lignification at the single cell level with improved sensitivity and resolution. This approach enables insights into the lignin composition in wall substructures, such as pits or in wall layers that are otherwise not distinguishable by classical LSCM. Our work emphasizes the importance of studying lignin formation in plant cell walls and demonstrates the potential of combining bioorthogonal chemistry and super-resolution microscopy techniques for studying biomolecules in living systems.

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“…Furthermore, as the improvements in obtainable optical resolution reach single nanometer precision, as it is the case for MINFLUX (minimal photon fluxes) − and MINSTED nanoscopy techniques, , the displacement of the fluorophore from the point of interest (referred to as linkage error) becomes a critical parameter. , Genetic code expansion (GCE) has emerged as a powerful platform for site-specific labeling of proteins with small organic fluorophores with minimal linkage error. , Through incorporation of unnatural amino acids (UAAs) with bioorthogonal reactivity to fluorescent probes, proteins of interest can be labeled in a fast and specific manner . The most prominent example is the combination of “clickable” UAAs and tetrazine-functionalized fluorophores for highly specific labeling by live-cell compatible strain-promoted inverse electron-demand Diels–Alder cycloaddition (SPIEDAC) reactions (Figure ).…”

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confidence: 99%

Bioorthogonal Caging-Group-Free Photoactivatable Probes for Minimal-Linkage-Error Nanoscopy

et al. 2023

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Here we describe highly compact, click compatible, and photoactivatable dyes for super-resolution fluorescence microscopy (nanoscopy). By combining the photoactivatable xanthone (PaX) core with a tetrazine group, we achieve minimally sized and highly sensitive molecular dyads for the selective labeling of unnatural amino acids introduced by genetic code expansion. We exploit the excited state quenching properties of the tetrazine group to attenuate the photoactivation rates of the PaX, and further reduce the overall fluorescence emission of the photogenerated fluorophore, providing two mechanisms of selectivity to reduce the offtarget signal. Coupled with MINFLUX nanoscopy, we employ our dyads in the minimal-linkage-error imaging of vimentin filaments, demonstrating molecular-scale precision in fluorophore positioning.

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Recent Advances in Fluorescence Imaging by Genetically Encoded Non-canonical Amino Acids

Cited by 19 publications

References 113 publications

Programmable Synthesis of Biobased Materials Using Cell‐Free Systems

Programmable Synthesis of Biobased Materials Using Cell‐Free Systems

Exploring Lignification Complexity in Plant Cell Walls with Airyscan Super-resolution Microscopy and Bioorthogonal Chemistry

Bioorthogonal Caging-Group-Free Photoactivatable Probes for Minimal-Linkage-Error Nanoscopy

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