A genetic circuit on a single DNA molecule as an autonomous dissipative nanodevice.

Realizing genetic circuits on single DNA molecules as self-encoded dissipative nanodevices is a major step toward miniaturization of autonomous biological systems. A circuit operating on a single DNA implies that genetically encoded proteins localize during coupled transcription-translation to DNA, but a single-molecule measurement demonstrating this has remained a challenge. Here, we use a genetically encoded fluorescent reporter system with improved temporal resolution and observe the synthesis of individual proteins tethered to a DNA molecule by transient complexes of RNA polymerase, messenger RNA, and ribosome. Against expectations in dilute cell-free conditions where equilibrium considerations favor dispersion, these nascent proteins linger long enough to regulate cascaded reactions on the same DNA. We rationally design a pulsatile genetic circuit by encoding an activator and repressor in feedback on the same DNA molecule. Driven by the local synthesis of only several proteins per hour and gene, the circuit dynamics exhibit enhanced variability between individual DNA molecules, and fluctuations with a broad power spectrum. Our results demonstrate that co-expressional localization, as a nonequilibrium process, facilitates single-DNA genetic circuits as dissipative nanodevices, with implications for nanobiotechnology applications and artificial cell design.

Systematic Tuning of Rhodamine Spirocyclization for Super-resolution Microscopy.

Rhodamines are the most important class of fluorophores for applications in live-cell fluorescence microscopy. This is mainly because rhodamines exist in a dynamic equilibrium between a fluorescent zwitterion and a nonfluorescent but cell-permeable spirocyclic form. Different imaging applications require different positions of this dynamic equilibrium, and an adjustment of the equilibrium poses a challenge for the design of suitable probes. We describe here how the conversion of the ortho-carboxy moiety of a given rhodamine into substituted acyl benzenesulfonamides and alkylamides permits the systematic tuning of the equilibrium of spirocyclization with unprecedented accuracy and over a large range. This allows one to transform the same rhodamine into either a highly fluorogenic and cell-permeable probe for live-cell-stimulated emission depletion (STED) microscopy or a spontaneously blinking dye for single-molecule localization microscopy (SMLM). We used this approach to generate differently colored probes optimized for different labeling systems and imaging applications.

Investigation of Lactones as Innovative Bio-Sourced Phase Change Materials for Latent Heat Storage.

In the presented work, five bio-based and bio-degradable cyclic esters, i.e. lactones, have been investigated as possible phase change materials for applications in latent heat storage systems. Commercial natural lactones such as ε-caprolactone and γ-valerolactone were easily purchased through chemical suppliers, while 1,2-campholide, oxa-adamantanone and dibenzochromen-6-one were synthesized through Baeyer-Villiger oxidation. The compounds were characterized with respect to attenuated total reflectance spectroscopy and gas chromatography coupled with mass spectroscopy, in order to confirm their chemical structures and identity. Subsequently, thermogravimetric analysis and differential scanning calorimetry were used to measure the phase change temperatures, enthalpies of fusion, degradation temperatures, as well to estimate the degree of supercooling. The lactones showed a wide range of phase change temperatures from -40 °C to 290 °C, making them a high interest for both low and high temperature latent heat storage applications, given the lack of organic phase change materials covering phase change temperature ranges below 0 °C and above 80 °C. However, low enthalpies of fusion, high degrees of supercooling and thermal degradations at low temperatures were registered for all samples, rendering them unsuitable as phase change materials.

The Furan Shuffling Hypothesis: A Biogenetic Proposal for Eremophilane Sesquiterpenoids.

Based on the structural similarities of the recently isolated eremophilane-type sesquiterpenoids microsphaeropsisin B and C and the iso-eremophilane periconianone C, a revised biogenetic hypothesis for C8-C11-connected iso-eremophilanes is presented and corroborated by strong experimental evidence. The first enantioselective total syntheses of microsphaeropsisin B and C were achieved starting from a known intermediate, whose synthesis was elaborated previously in the total synthesis of periconianone A, and in a total of 15 steps starting from γ-hydroxy carvone. Mild reaction conditions for the subsequent α-ketol rearrangement not only resulted in the herein proposed conversion of microsphaeropsisin B into periconianone C, but also in the conversion of microsphaeropsisin C into 4-epi-periconianone C.