Isonitrile induced bioorthogonal activation of fluorophores and mutually orthogonal cleavage in live cells

2021 ◽  
Author(s):  
Xiaoyang Zhang ◽  
Hui Xu ◽  
Jie Li ◽  
Dunyan Su ◽  
Wuyu Mao ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Live Cells ◽  
Release Reaction ◽  
Tert Butyl

Fluorophores with different emission wavelengths were efficiently quenched by tert-butyl terminated tetrazylmethyl group and activated by isonitrile-tetrazine click-to-release reaction. Nucleic acid templated chemistry significantly accelerated this bioorthogonal cleavage. Moreover, two...

scholarly journals Nucleic‐Acid Structures as Intracellular Probes for Live Cells

2019 ◽  
Vol 32 (13) ◽  
pp. 1901743 ◽  
Author(s):  
Devleena Samanta ◽  
Sasha B. Ebrahimi ◽  
Chad A. Mirkin
Keyword(s):  
Nucleic Acid ◽  
Live Cells ◽  
Nucleic Acid Structures

scholarly journals Controlled exchange of protein and nucleic acid signals from and between synthetic minimal cells

2022 ◽  
Author(s):  
Joseph M Heili ◽  
Kaitlin Stokes ◽  
Nathaniel J Gaut ◽  
Christopher Deich ◽  
Jose Gomez-Garcia ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Rna Binding ◽  
Rna Binding Proteins ◽  
Live Cells ◽  
Minimal Cell ◽  
Functional Protein ◽  
Vesicle Membrane ◽  
Synthetic Cell ◽  
Peptide Tags ◽  
Minimal Cells

Synthetic minimal cells are a class of small liposome bioreactors that have some, but not all functions of live cells. Here, we report a critical step towards the development of a bottom-up minimal cell: cellular export of functional protein and RNA products. We used cell penetrating peptide tags to translocate payloads across a synthetic cell vesicle membrane. We demonstrated efficient transport of active enzymes, and transport of nucleic acid payloads by RNA binding proteins. We investigated influence of a concentration gradient alongside other factors on the efficiency of the translocation, and we show a method to increase product accumulation in one location. We demonstrate the use of this technology to engineer molecular communication between different populations of synthetic cells, to exchange protein and nucleic acid signals. The synthetic minimal cell production and export of proteins or nucleic acids allows experimental designs that approach the complexity and relevancy of natural biological systems.

Nucleic acid related compounds. 34. Non-aqueous diazotization with tert-butyl nitrite. Introduction of fluorine, chlorine, and bromine at C-2 of purine nucleosides

1981 ◽  
Vol 59 (17) ◽  
pp. 2608-2611 ◽  
Author(s):  
Morris J. Robins ◽  
Bogdan Uznański
Keyword(s):  
Nucleic Acid ◽  
Hydrogen Fluoride ◽  
Antimony Trichloride ◽  
Anhydrous Ammonia ◽  
Pyridine Hydrochloride ◽  
Tert Butyl ◽  
Anhydrous Hydrogen Fluoride ◽  
Related Compounds

Treatment of 2,6-diamino-9-(2,3,5-tri-O-acetyl-β-D-ribofuranosyl)purine (1c) with tert-butyl nitrite (TBN) in 60% anhydrous hydrogen fluoride/pyridine at −20 °C gave 2-fluoroadenosine triacetate (2) in 48% yield. Analogous treatment of the acetylated 2-amino-6-fluoropurine nucleoside (1a) gave the 2,6-difluoro compound (3a) in 63% yield. Anhydrous ammonia in 1,2-dimethoxy-ethane converted 3a → 2 without ammonolysis of the acetate groups. The overall conversion of 1a → 3a → 2 without isolation of 3a proceeded in 80% yield. Similar diazotization of the 2-amino-6-chloro nucleoside (1b) in HF/pyridine gave the 2-fluoro-6-chloro analogue (3b) in 89% yield. Treatment of 1b with TBN and pyridine hydrochloride in dichloromethane (77%) or TBN and antimony trichloride in 1,2-dichloroethane (84%) gave the 2,6-dichloropurine nucleoside (4b). The latter conditions converted 1a to the 2-chloro-6-fluoro product (4a) in 77% yield. Conversions of 1a and 1b to the 2-bromo-6-fluoro (5a, 67%) and 2-bromo-6-chloro (5b, 78%) analogues were effected using TBN and antimony tribromide in dibromomethane.

scholarly journals A smart ZnO@polydopamine-nucleic acid nanosystem for ultrasensitive live cell mRNA imaging by the target-triggered intracellular self-assembly of active DNAzyme nanostructures

2017 ◽  
Vol 8 (4) ◽  
pp. 2832-2840 ◽  
Author(s):  
Dinggeng He ◽  
Xing He ◽  
Xue Yang ◽  
Hung-Wing Li
Keyword(s):  
Nucleic Acid ◽  
Self Assembly ◽  
Live Cell ◽  
Live Cells

We have developed a smart ZnO@PDA-hpDNAs nanosystem carrying four functional hpDNA strands that can realize the target-triggered self-assembly of wire-shaped active DNAzyme nanostructures via HCR in live cells.

scholarly journals Nucleic acid dyes for detection of apoptosis in live cells

Cytometry ◽  
1995 ◽  
Vol 21 (3) ◽  
pp. 265-274 ◽  
Author(s):  
Tom Frey
Keyword(s):  
Nucleic Acid ◽  
Live Cells ◽  
Acid Dyes

scholarly journals Nucleic acid-selective light-up fluorescent biosensors for ratiometric two-photon imaging of the viscosity of live cells and tissues

2016 ◽  
Vol 7 (3) ◽  
pp. 2257-2263 ◽  
Author(s):  
Dandan Li ◽  
Xiaohe Tian ◽  
Aidong Wang ◽  
Lijuan Guan ◽  
Jun Zheng ◽  
...  
Keyword(s):  
Nucleic Acid ◽  
Nuclear Dna ◽  
Water Soluble ◽  
Live Cells ◽  
Fluorescent Biosensors ◽  
Two Photon ◽  
High Affinity ◽  
Dna And Rna ◽  
Viscosity Sensors ◽  
Photon Imaging

Two water-soluble TPEF probes which can be applied as ratiometric viscosity sensors were shown to bind to nuclear DNA and RNA in the nucleolus and cytoplasm with high affinity, respectively.

scholarly journals Evaluating the Flow-Cytometric Nucleic Acid Double-Staining Protocol in Realistic Situations of Planktonic Bacterial Death

2008 ◽  
Vol 74 (6) ◽  
pp. 1767-1779 ◽  
Author(s):  
Tania Falcioni ◽  
Stefano Papa ◽  
Josep M. Gasol
Keyword(s):  
Nucleic Acid ◽  
Extreme Environments ◽  
Membrane Integrity ◽  
High Capacity ◽  
Live Cells ◽  
Leucine Incorporation ◽  
Double Staining ◽  
Tetrazolium Chloride ◽  
Physiological Diversity ◽  
Staining Protocol

ABSTRACT Since heterotrophic prokaryotes play an important biogeochemical role in aquatic ecosystems and have a high capacity to survive in extreme environments, easy-to-perform protocols that probe their physiological states and the effects of environmental variables on those states are highly desired. Some methodologies combine a general nucleic acid stain with a membrane integrity probe. We calibrated one of these, the nucleic acid double-staining (NADS) protocol (G. Grégori, S. Citterio, A. Ghiani, M. Labra, S. Sgorbati, S. Brown, and M. Denis, Appl. Environ. Microbiol. 67:4662-4670, 2001), determining the optimal stain concentrations in seawater and the response to conditions that generate prokaryote death (such as heat) and to conditions that are known to produce death in plankton, such as nutrient limitation or flagellate grazing. The protocol was validated by comparison to two methods used to detect viability: active respiration by 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) and incorporation of tritiated leucine. We show that concentrations in the range of 5 to 20 μg ml−1 of propidium iodide, simultaneous to a 10× concentration of Sybr green I, are best for detecting two separated populations of “live” (green cells) and “dead” (red cells) organisms. During exposure to heat and UVC, we observed that the number of live cells declined concurrently with that of actively respiring cells (CTC positive) and with total leucine incorporation. In seawater mesocosms, the NADS protocol allowed detection of bacterioplankton starvation-related death and flagellate predation. The protocol was also tested in deep profiles in the northwest Atlantic, demonstrating its potential for routine characterization of this fraction of the physiological diversity of marine heterotrophic prokaryotic plankton.

scholarly journals Fluorescent Probes for Nucleic Acid Visualization in Fixed and Live Cells

Molecules ◽  
2013 ◽  
Vol 18 (12) ◽  
pp. 15357-15397 ◽  
Author(s):  
Alexandre Boutorine ◽  
Darya Novopashina ◽  
Olga Krasheninina ◽  
Karine Nozeret ◽  
Alya Venyaminova
Keyword(s):  
Nucleic Acid ◽  
Fluorescent Probes ◽  
Live Cells
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