Here, we show how dynamic nuclear polarization (DNP) NMR spectroscopy experiments permit the atomic level structural characterization of loaded and empty lipid nanoparticles (LNPs). The LNPs used here were synthesized by the microfluidic mixing technique and are composed of ionizable cationic lipid (DLin-MC3-DMA), a phospholipid (distearoylphosphatidylcholine, DSPC), cholesterol, and poly(ethylene glycol) (PEG) (dimyristoyl phosphatidyl ethanolamine (DMPE)-PEG 2000), as well as encapsulated cargoes that are either phosphorothioated siRNA (50 or 100%) or mRNA. We show that LNPs form physically stable complexes with bioactive drug siRNA for a period of 94 days. Relayed DNP experiments are performed to study H-H spin diffusion and to determine the spatial location of the various components of the LNP by studying the average enhancement factors as a function of polarization time. We observe a striking feature of LNPs in the presence and in the absence of encapsulating siRNA or mRNA by comparing our experimental results to numerical spin-diffusion modeling. We observe that LNPs form a layered structure, and we detect that DSPC and DMPE-PEG 2000 lipids form a surface rich layer in the presence (or absence) of the cargoes and that the cholesterol and ionizable cationic lipid are embedded in the core. Furthermore, relayed DNP P solid-state NMR experiments allow the location of the cargo encapsulated in the LNPs to be determined. On the basis of the results, we propose a new structural model for the LNPs that features a homogeneous core with a tendency for layering of DSPC and DMPE-PEG at the surface.

No abstract

The CRISPR-Cas9 system has increased the speed and precision of genetic editing in cells and animals. However, model generation for drug development is still expensive and time-consuming, demanding more target flexibility and faster turnaround times with high reproducibility. The generation of a tightly controlled ObLiGaRe doxycycline inducible SpCas9 (ODInCas9) transgene and its use in targeted ObLiGaRe results in functional integration into both human and mouse cells culminating in the generation of the ODInCas9 mouse. Genomic editing can be performed in cells of various tissue origins without any detectable gene editing in the absence of doxycycline. Somatic in vivo editing can model non-small cell lung cancer (NSCLC) adenocarcinomas, enabling treatment studies to validate the efficacy of candidate drugs. The ODInCas9 mouse allows robust and tunable genome editing granting flexibility, speed and uniformity at less cost, leading to high throughput and practical preclinical in vivo therapeutic testing.

<p><span>The plate boundary between the American and Eurasian plates runs in southwest Iceland along a 5-10 km wide seismicity zone on the Reykjanes Peninsula. There, tectonic spreading events take place as continuous seismic release and seismic episodes (swarms and individual large events) with recurrence interval of about 40 years and volcanic episodes at intervals of 800-1000 years. The crust in Reykjanes is, therefore, particularly thin and hot and geothermal energy is currently harnesses in two areas on the western part of the peninsula in Reykjanes and Svartsengi.</span></p><p><span>Since January 2020, earthquake swarms with larger events up to M5.6 have been occurring frequently over the entire Reykjanes Peninsula, accompanied by unusual uplift (up to 12 cm) and subsidence cycles in the Svartsengi-Eldvörp fissure swarm. This raises the question whether we might be at the beginning of a new volcanic episode. In order to classify such processes at an early stage, multidisciplinary geophysical measurements are particularly valuable.</span></p><p>The Icelandic Meteorological Office (IMO), University of Iceland as well as <span>ISOR and several partners responded immediately after the unrest began. As soon as January 2020, GFZ proposed a rapid response field campaign (MAGIC: MultidisciplinAry imaGIng and Characterization of the magma/fluid reservoir beneath Svartsengi). Only one week after the uplift start and first earthquake swarm, we connected a Distributed Acoustic Sensing interrogator to a 21 km long telecommunication fibre optic cable which crosses the uplift and swarm area. In addition, while we complied to strict constraints due to the Covid-19 pandemic, the rapid response activities comprised deployment of several additional sensors including broadband seismology, rotational seismology and we performed repeated surveys including gas-, gravity-, </span><span>electromagnetic</span><span>-, airborne and ground magnetic- measurements. </span></p><p><span>We present preliminary results from various techniques and discuss their role in discriminating different scenarios aiming at explaining the magma-tectonic unrest phase. In particular, we analyze how the combination of airborne snapshots of ground morphology can be combined with the high temporal and spatial resolution deformation fields along the fibre optic cable. </span></p>

The CRISPR-Cas9 system has increased the speed and precision of genetic editing in cells and animals. However, model generation for drug development is still expensive and time-consuming, demanding more target flexibility and faster turnaround times with high reproducibility. We have generated a tightly controlled ObLiGaRe doxycycline inducible SpCas9 (ODInCas9) transgene. Targeted ObLiGaRe resulted in functional integration into both human and mouse cells culminating in the generation of the ODInCas9 mouse. Genomic editing can be performed in cells of various tissue origins without any detectable gene editing in the absence of doxycycline. Somatic in vivo editing can model non-small cell lung cancer (NSCLC) adenocarcinomas, enabling treatment studies to validate the efficacy of candidate drugs. The ODInCas9 mouse can be utilized for robust and tunable genome editing allowing for flexibility, speed and uniformity at reduced cost, leading to high throughput and practical preclinical in vivo therapeutic testing.