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Abstract Naturally generated lipid nanoparticles termed extracellular vesicles (EVs) hold significant promise as engineerable therapeutic delivery vehicles. However, active loading of protein cargo into EVs in a manner that is useful for delivery remains a challenge. Here, we demonstrate that by rationally designing proteins to traffic to the plasma membrane and associate with lipid rafts, we can enhance loading of protein cargo into EVs for a set of structurally diverse transmembrane and peripheral membrane proteins. We then demonstrate the capacity of select lipid tags to mediate increased EV loading and functional delivery of an engineered transcription factor to modulate gene expression in target cells. We envision that this technology could be leveraged to develop new EV-based therapeutics that deliver a wide array of macromolecular cargo. 

Abstract The organization of membrane proteins between and within membrane-bound compartments is critical to cellular function. Yet we lack approaches to regulate this organization in a range of membrane-based materials, such as engineered cells, exosomes, and liposomes. Uncovering and leveraging biophysical drivers of membrane protein organization to design membrane systems could greatly enhance the functionality of these materials. Towards this goal, we use de novo protein design, molecular dynamic simulations, and cell-free systems to explore how membrane-protein hydrophobic mismatch could be used to tune protein cotranslational integration and organization in synthetic lipid membranes. We find that membranes must deform to accommodate membrane-protein hydrophobic mismatch, which reduces the expression and co-translational insertion of membrane proteins into synthetic membranes. We use this principle to sort proteins both between and within membranes, thereby achieving one-pot assembly of vesicles with distinct functions and controlled split-protein assembly, respectively. Our results shed light on protein organization in biological membranes and provide a framework to design self-organizing membrane-based materials with applications such as artificial cells, biosensors, and therapeutic nanoparticles. 

Palmer Deep sediment cores are used to produce the first high-resolution, continuous late Pleistocene to Holocene time-series from the Antarctic marine system. The sedimentary record is dated using accelerator mass spectrometer radiocarbon methods on acid insoluble organic matter and foraminiferal calcite. Fifty-four radiocarbon analyses are utilized in the dating which provides a calibrated timescale back to 13 ka BP. Reliability of resultant ages on organic matter is assured because duplicates produce a standard deviation from the surface age of less than laboratory error (i.e., ±50 years). In addition, surface organic matter ages at the site are in excellent agreement with living calcite ages at the accepted reservoir age of 1260 years for the Antarctic Peninsula. Spectral analyses of the magnetic susceptibility record against the age model reveal unusually strong periodicity in the 400,–200 and 50-70 year frequency bands, similar to other high-resolution records from the Holocene but, so far, unique for the circum-Antarctic. Here we show that comparison to icecore records of specific climatic events (e.g., the ’Little Ice Age‘, Neoglacial, Hypsithermal, and the Bølling/Allerød to Younger Dryas transition) provides improved focus upon the relative timing of atmosphere/ocean changes between the northern anid southern high latitudes.