Photoinducible Azobenzene trimethylammonium bromide (AzoTAB)-mediated giant vesicle fusion compatible with synthetic protein translation reactions

Xu, Bo-Ying; Ding, Jinquan; Caliari, Adriano; Lu, Nan; Han, Fuhai; Xia, Yang; Xu, Jian; Yomo, Tetsuya

doi:10.1016/j.bbrc.2022.06.035

Cited by 4 publications

(3 citation statements)

References 30 publications

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“…Some examples are shown in Scheme 1. cis–trans Isomerism causes the photochromism of azobenzenes 1 , 2 and structural isomerism results in photochromism of spiropyrans 2 , 3 furylfulgides 3 4 and bis(thienyl)ethenes 4 . 1 Whereas the former reaction is a 6- endo -trig ring-closure under neutralization of the partial charges of the push–pull-chromophor, the two latter mentioned reactions are formal 6π-electrocyclizations to form ring-closed isomers.…”

Section: Introductionmentioning

confidence: 99%

Bis(thienyl)ethenes with α-methoxymethyl groups. Syntheses, spectroscopic Hammett plots, and stabilities in PMMA films

Weingartz,

Nagorny,

Adams

et al. 2023

RSC Adv.

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Section: Introductionmentioning

confidence: 99%

Bis(thienyl)ethenes with α-methoxymethyl groups. Syntheses, spectroscopic Hammett plots, and stabilities in PMMA films

Weingartz,

Nagorny,

Adams

et al. 2023

RSC Adv.

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“…Flow cytometry was one of the first techniques applied to this end, but it is limited due to its lack of spatial resolution [27][28][29]. Imaging flow cytometry (IFC) provides significant improvements through the collection of widefield fluorescence imaging data for thousands of objects in a flow chamber [14,30,31]. Although valuable, IFC is not yet widely available, nor does it allow data acquisition for GUV in their native state and under controlled conditions.…”

Section: Introductionmentioning

confidence: 99%

“…Previous techniques approached the problem using fluorogenic reactions or protein expression, which are triggered following content mixing. Exchange of lipidic components has been monitored by changes in fluorescent lipid emission due to Förster resonance energy transfer [10,31]. Taking advantage of the size of GUV, microscopy, or IFC to confirm fusion, provides a convenient alternative assay based on the colocalization of two fluorophores, either encapsulated in the lumen (content markers), or localized on the membrane (lipid markers) of separate populations of GUV.…”

Section: Introductionmentioning

confidence: 99%

Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis

Caliari

Hanczyc

Imai

et al. 2023

IJMS

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Artificial cells are based on dynamic compartmentalized systems. Thus, remodeling of membrane-bound systems, such as giant unilamellar vesicles, is finding applications beyond biological studies, to engineer cell-mimicking structures. Giant unilamellar vesicle fusion is rapidly becoming an essential experimental step as artificial cells gain prominence in synthetic biology. Several techniques have been developed to accomplish this step, with varying efficiency and selectivity. To date, characterization of vesicle fusion has relied on small samples of giant vesicles, examined either manually or by fluorometric assays on suspensions of small and large unilamellar vesicles. Automation of the detection and characterization of fusion products is now necessary for the screening and optimization of these fusion protocols. To this end, we implemented a fusion assay based on fluorophore colocalization on the membranes and in the lumen of vesicles. Fluorescence colocalization was evaluated within single compartments by image segmentation with minimal user input, allowing the application of the technique to high-throughput screenings. After detection, statistical information on vesicle fluorescence and morphological properties can be summarized and visualized, assessing lipid and content transfer for each object by the correlation coefficient of different fluorescence channels. Using this tool, we report and characterize the unexpected fusogenic activity of sodium chloride on phosphatidylcholine giant vesicles. Lipid transfer in most of the vesicles could be detected after 20 h of incubation, while content exchange only occurred with additional stimuli in around 8% of vesicles.

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Correlation between Absorption and Substitution of Photochromic 1,2‐Bis(thienyl)ethenes (BTEs) Using Modified Spectroscopic Hammett Equations

et al. 2022

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Series of photochromic 1,2-bis(thienyl)ethenes possessing perfluorocyclopentene backbones, either hydrogen or methyl groups at the β-positions of the thiophenes, and a variety of substituents in their α'-positions were prepared, which cover the range from electron-donating to electron-withdrawing (Me, À CH 2 OH, À OTBS, À TMS, À Br, 1,3-dioxan-2-yl, pyridin-4-yl, À CH 2 OH, À COOH). As a linear free energy relationship the spectroscopic Hammett equation gives fair to excellent fits to the excitation energy of the absorption maxima of the ring-opened as well as the ring-closed forms of the BTEs, when Hammett substituent constants σ p were replaced by Brown's modified substituent constants σ p + and σ p À . Vice versa, hitherto unknown Hammett-Brown substituent constants can be estimated from the UV spectra. Furthermore, we compared the experimentally measured absorption maxima with values which we calculated by three different methods (DFT STEOM-DLPNO-CCSD/def2-TZVPP, TD-DFT ωB97X-D3/6-31G*, TD-DFT ωB97X-D3/6-311 + + G**).

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Photoinducible Azobenzene trimethylammonium bromide (AzoTAB)-mediated giant vesicle fusion compatible with synthetic protein translation reactions

Cited by 4 publications

References 30 publications

Bis(thienyl)ethenes with α-methoxymethyl groups. Syntheses, spectroscopic Hammett plots, and stabilities in PMMA films

Bis(thienyl)ethenes with α-methoxymethyl groups. Syntheses, spectroscopic Hammett plots, and stabilities in PMMA films

Quantification of Giant Unilamellar Vesicle Fusion Products by High-Throughput Image Analysis

Correlation between Absorption and Substitution of Photochromic 1,2‐Bis(thienyl)ethenes (BTEs) Using Modified Spectroscopic Hammett Equations

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