An improved toolkit of gateway- and gibson assembly-compatible vectors for protoplast transfection and agrobacterium-mediated plant transformation DOI Creative Commons
Jeffrey R. Allen, Michael Flanagan, Sunita Pathak

et al.

BMC Research Notes, Journal Year: 2025, Volume and Issue: 18(1)

Published: April 8, 2025

Understanding the regulation and function of plant genes is essential for addressing challenges faced by modern agriculture. Plant transformation, in conjunction with fluorescence microscopy, offers a powerful approach to investigate dynamic behavior proteins they encode. We previously developed set Gateway-compatible tissue-specific transformation vectors. In this paper we aim expand toolkit vectors available Agrobacterium-mediated protoplast transfection. Here, introduce new introducing additional fluorophores create pJRA vector series. Additionally, pLCS series vectors, modular Gateway- Gibson assembly-compatible designed All described are from Addgene serve as resource research community.

Language: Английский

Bioorthogonal Chemistry and Its Applications DOI Creative Commons
Robert E. Bird,

S. A. Lemmel,

Xiang Yu

et al.

Bioconjugate Chemistry, Journal Year: 2021, Volume and Issue: 32(12), P. 2457 - 2479

Published: Nov. 30, 2021

Bioorthogonal chemistry is a set of methods using the non-native functional groups to explore and understand biology in living organisms. In this review, we summarize most common reactions used bioorthogonal methods, their relative advantages disadvantages, frequency occurrence published literature. We also briefly discuss some less but potentially useful methods. then analyze bioorthogonal-related publications CAS Content Collection determine how often different types biomolecules such as proteins, carbohydrates, glycans, lipids have been studied chemistry. The prevalent biological chemical for attaching these are elaborated. publication volume related applications Collection. use imaging, identifying, characterizing delivering drugs treat disease discussed at length. surface attachment proteins modified carbohydrates noted. Finally, state art its current limitations promise future productive biology.

Language: Английский

Citations

245

Focal adhesion dynamics in cellular function and disease DOI
Yasaswi Gayatri Mishra, Bramanandam Manavathi

Cellular Signalling, Journal Year: 2021, Volume and Issue: 85, P. 110046 - 110046

Published: May 15, 2021

Language: Английский

Citations

125

Tissue clearing DOI
Douglas S. Richardson, Webster Guan, Katsuhiko Matsumoto

et al.

Nature Reviews Methods Primers, Journal Year: 2021, Volume and Issue: 1(1)

Published: Dec. 16, 2021

Language: Английский

Citations

105

Exchangeable HaloTag Ligands for Super-Resolution Fluorescence Microscopy DOI Creative Commons
Julian Kompa, Jorick J. Bruins, Marius Glogger

et al.

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(5), P. 3075 - 3083

Published: Jan. 30, 2023

The specific and covalent labeling of the protein HaloTag with fluorescent probes in living cells makes it a powerful tool for bioimaging. However, irreversible attachment probe to precludes imaging applications that require transient binding comes risk photobleaching. Here, we introduce exchangeable ligands fluorescence (xHTLs) reversibly bind can be coupled rhodamines different colors. In stimulated emission depletion (STED) microscopy, exchange xHTLs allows reduced photobleaching as compared labeling. Transient fluorogenic fusion proteins enables points accumulation nanoscale topography (PAINT) MINFLUX microscopy. We furthermore pairs mutants dual-color PAINT STED thus open up new possibilities across microscopy platforms widely used approach.

Language: Английский

Citations

83

Challenges in the use of sortase and other peptide ligases for site-specific protein modification DOI Creative Commons
Holly E. Morgan, W. Bruce Turnbull, Michael E. Webb

et al.

Chemical Society Reviews, Journal Year: 2022, Volume and Issue: 51(10), P. 4121 - 4145

Published: Jan. 1, 2022

We highlight chemical and biochemical strategies taken to optimise peptide protein modification using ligases.

Language: Английский

Citations

77

Xanthene, cyanine, oxazine and BODIPY: the four pillars of the fluorophore empire for super-resolution bioimaging DOI
Soham Samanta, Kaitao Lai,

Feihu Wu

et al.

Chemical Society Reviews, Journal Year: 2023, Volume and Issue: 52(20), P. 7197 - 7261

Published: Jan. 1, 2023

In the realm of biological research, invention super-resolution microscopy (SRM) has enabled visualization ultrafine sub-cellular structures and their functions in live cells at nano-scale level, beyond diffraction limit, which opened up a new window for advanced biomedical studies to unravel complex unknown details physiological disorders level with unprecedented resolution clarity. However, most SRM techniques are highly reliant on personalized special photophysical features fluorophores. recent times, there been an surge development robust fluorophore systems various imaging techniques. To date, xanthene, cyanine, oxazine BODIPY cores have authoritatively utilized as basic units small-molecule-based organic fluorescent probe designing strategies owing excellent characteristics easy synthetic acquiescence. Since future next-generation will be decided by availability probes these four building blocks play important role progressive design, is urgent need review advancements fluorophores different methods based dye cores. This article not only includes comprehensive discussion about developments emphasis effective integration into cell bio-imaging applications but also critically evaluates background each highlight merits demerits towards developing newer SRM.

Language: Английский

Citations

63

Hybrid Small-Molecule/Protein Fluorescent Probes DOI
Masafumi Minoshima, Shahi Imam Reja,

Ryu Hashimoto

et al.

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(10), P. 6198 - 6270

Published: May 8, 2024

Hybrid small-molecule/protein fluorescent probes are powerful tools for visualizing protein localization and function in living cells. These hybrid constructed by diverse site-specific chemical labeling approaches through reactions to exogenous peptide/small tags, enzymatic post-translational modifications, bioorthogonal genetically incorporated unnatural amino acids, ligand-directed reactions. The employed imaging trafficking, conformational changes, bioanalytes surrounding proteins. In addition, facilitate visualization of dynamics at the single-molecule level defined structure with super-resolution imaging. this review, we discuss development bioimaging applications based on hybrids.

Language: Английский

Citations

21

A Comprehensive Guide to Enzyme Immobilization: All You Need to Know DOI Creative Commons
Marina Simona Robescu, Teodora Bavaro

Molecules, Journal Year: 2025, Volume and Issue: 30(4), P. 939 - 939

Published: Feb. 18, 2025

Enzyme immobilization plays a critical role in enhancing the efficiency and sustainability of biocatalysis, addressing key challenges such as limited enzyme stability, short shelf life, difficulties recovery recycling, which are pivotal for green chemistry industrial applications. Classical approaches, including adsorption, entrapment, encapsulation, covalent bonding, well advanced site-specific methods that integrate engineering bio-orthogonal chemistry, were discussed. These techniques enable precise control over orientation interaction with carriers, optimizing catalytic activity reusability. Key findings highlight impact on improving performance under various operational conditions its reducing process costs through enhanced stability recyclability. The review presents numerous practical applications immobilized enzymes, their use pharmaceutical industry drug synthesis, food sector dairy processing, environmental biotechnology wastewater treatment dye degradation. Despite significant advantages, loss due to conformational changes mass transfer limitations remain, necessitating tailored protocols specific integration modern biotechnological advancements, site-directed mutagenesis recombinant DNA technology, offers promising pathway developing robust, efficient, sustainable biocatalytic systems. This comprehensive guide aims support researchers industries selecting diverse pharmaceuticals, fine chemicals.

Language: Английский

Citations

2

A brain atlas of synapse protein lifetime across the mouse lifespan DOI Creative Commons
Edita Bulovaite, Zhen Qiu,

Maximilian Kratschke

et al.

Neuron, Journal Year: 2022, Volume and Issue: 110(24), P. 4057 - 4073.e8

Published: Oct. 5, 2022

The lifetime of proteins in synapses is important for their signaling, maintenance, and remodeling, memory duration. We quantified the endogenous PSD95, an abundant postsynaptic protein excitatory synapses, at single-synapse resolution across mouse brain lifespan, generating Protein Lifetime Synaptome Atlas. Excitatory have a wide range PSD95 lifetimes extending from hours to several months, with distinct spatial distributions dendrites, neurons, regions. Synapses short are enriched young animals regions controlling innate behaviors, whereas long accumulate during development, cortex CA1 where memories stored, preferentially preserved old age. Synapse increases throughout model autism schizophrenia. adds further layer synapse diversity enriches prevailing concepts aging, disease.

Language: Английский

Citations

66

HaloTag‐Based Reporters for Fluorescence Imaging and Biosensing DOI Creative Commons
Alexander Cook, Franziska Walterspiel, Claire Deo

et al.

ChemBioChem, Journal Year: 2023, Volume and Issue: 24(12)

Published: Feb. 23, 2023

Abstract Visualizing the structure and dynamics of biomolecules is critical to understand biological function, requires methods fluorescently label targets interest in their cellular context. Self‐labelling proteins, which combine a genetically encoded tag with small‐molecule fluorophore, have attracted considerable attention for this purpose, as they can overcome limitations fluorescent proteins. Among them, HaloTag protein most broadly used, showing fast specific labelling small, easy functionalize cell‐permeant ligand. Synthetic chemistry engineering provided portfolio powerful imaging tools exploiting HaloTag, along general optimize adapt them applications. Here, we provide an overview reporters based on biosensing, highlighting strategies

Language: Английский

Citations

34