National Technology & Engineering Solutions of Sandia, LLC

Sandia Corporation

Sandia National Laboratories

Other

Arizona State University

Sandia Naitonal Laboratories

Sandia National Corporation

Joseph S Schoeniger

Phillip Harris Paul

Don Wesley Arnold

David W Neyer

Brooke Nicole Harmon

Edwin A Saada

David J Rakestraw

Yooli Kim Light

Kenneth R Hencken

Oscar Negrete

Luke Schoeniger

Anup Kumar Singh

Eric Bryant Cummings

James S Brennan

Ryan Wesley Davis

Stanley H Kravitz

Robert C Hughes

Richard Janek

Nancy Yuan Yang

Antonio J Ricco

Karl Wally

Hung Pin Kao

Gregory F Cardinale

Kenneth Sale

Arul Mozhy Varman

Mary Bao Tran-Gyamfi

Michael G Garguilo

Annette Estelle LaBauve

Aditya Pandharinath Sarnaik

Kyle Jeffrey Seamon

Gary Kruppa

Malin M Young

Albert W Flounders

Peter Anderson

Muhammad Faisal

Masood Zia Hadi

Specific and broad-spectrum inhibitory peptides to CRISPR Cas9 variants (SpyCas9, SauCas9, and CjeCas9) are disclosed. A method of making and using these peptides with therapeutic CRISPR Cas9 (to improve desired targeting) or against harmful CRISPR Cas9 components (already active in a patient) is also disclosed. The peptides combined with a delivery system is also disclosed.

Peptide inhibitors of CRISPR-Cas9

The present invention relates, in part, to methods for detecting nuclease activity, such as the activity of Cas nucleases. Also described herein are compositions for conducting assays, as well as methods for conducting assays in the presence of test compounds.

CRISPR/Cas activity assays and compositions thereof

An in vitro assay was designed to measure the activity of the alphavirus non-structural protein 2 (nsP2), which is the viral protease and is required for viral replication. By taking advantage of fluorescence-resonance energy transfer between two proteins, a protease cleavage assay was generated. This was utilized for high-throughput screening of 40,000 small molecules. Inhibitors were validated using cell-based assays to measure alphavirus infection and cytotoxicity. Certain compounds were then characterized for anti-viral efficacy in various cell lines in numerous assays. Compounds were tested against Chikungunya virus, Venezuelan Equine Encephalitis virus, Rift Valley Fever virus, and Zika virus. Three compounds (compounds I, II, and III) showed pan-alphavirus anti-viral efficacy at concentrations that did not result in cell toxicity. An additional compound, structure IV, showed broad spectrum inhibition of all viruses tested. Pharmaceutical preparations and methods of treatment including these compounds are provided herein.

Small molecule inhibitors for treatment of alpha viruses

A method of improving a ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCo) to have a higher protein score is disclosed. The method includes the steps of: making a modified RbcL of the RuBisCo, by, on an RbcL unit of the RuBisCo, either substituting Met for Leu, Phe, Val, or Ile or combinations thereof; substituting Lys for Arg, Thr, or His or combinations thereof; or both of these substitutions. The modified RbcL consequently modifies the RuBisCo and is added to a biomass host where it is stable for homologous recombination. Plastid and nucleus integration was observed. Example RbcL sequences are disclosed with the desirable substitutions. The improved RuBisCo can be used as an improved proteinaceous food source for humans and animals.

Engineering RuBisCo for food safety

A construct includes a core comprising an external surface and a plurality of pores, a cargo disposed in a pore of the plurality of pores, the cargo comprising a CRISPR Cas9 component or a nucleic acid sequence encoding a CRISPR Cas9 component; and a coating coupled to the core, wherein the coating comprises a cationic lipid, a pegylated lipid, a zwitterionic lipid, and a sterol. The coating may comprise a molar ratio of about 1 cationic lipid to 1 zwitterionic lipid to 0.9 sterol to 0.15 PEGylated lipid, wherein each molar ratio optionally varies by about plus or minus 10%. A method of treatment is also provided.

Lipid composition for the delivery of therapeutic cargos

A method including identifying one or more structures in adjacent sequences of one or more proteins that can be mimicked by a peptide; and assembling a peptide that mimics the one or more secondary structures. A method including assembling epitopes of linked pairs of discontiguous peptide subsequences that mimic one or more structures in sequences of a protein; and assaying antibodies against the epitopes.

Linear peptides that mimic structural epitopes for antibody assays

The present invention relates to nucleic acids, peptides, vectors, cells, and plants useful in the production of biofuels. In certain embodiments, the invention relates to nucleic acid sequences and peptides from extremophile organisms, such as SSO1949 and Ce1A, that are useful for hydrolyzing plant cell wall materials. In further embodiments, the invention relates to modified versions of such sequences that have been optimized for production in one or both of monocot and dicot plants. In other embodiments, the invention provides for targeting peptide production or activity to a certain location within the cell or organism, such as the apoplast. In further embodiments, the invention relates to transformed cells or plants. In additional embodiments, the invention relates to methods of producing biofuel utilizing such nucleic acids, peptides, targeting sequences, vectors, cells, and/or plants.

Methods and materials for deconstruction of biomass for biofuels production

The present invention relates to novel methods of determining the sequence and structure of proteins. Specifically, the present invention allows for the analysis of intact proteins within a mass spectrometer. Therefore, preparatory separations need not be performed prior to introducing a protein sample into the mass spectrometer. Also disclosed herein are new instrumental developments for enhancing the signal from the desired modified proteins, methods for producing controlled protein fragments in the mass spectrometer, eliminating complex microseparations, and protein preparatory chemical steps necessary for cross-linking based protein structure determination. Additionally, the preferred method of the present invention involves the determination of protein structures utilizing a top-down analysis of protein structures to search for covalent modifications. In the preferred method, intact proteins are ionized and fragmented within the mass spectrometer.

Structural determination of intact proteins using mass spectrometry

The present invention discloses an electrochemical device for detecting single particles, and methods for using such a device to achieve high sensitivity for detecting particles such as bacteria, viruses, aggregates, immuno-complexes, molecules, or ionic species. The device provides for affinity-based electrochemical detection of particles with single-particle sensitivity. The disclosed device and methods are based on microelectrodes with surface-attached, affinity ligands (e.g., antibodies, combinatorial peptides, glycolipids) that bind selectively to some target particle species. The electrodes electrolyze chemical species present in the particle-containing solution, and particle interaction with a sensor element modulates its electrolytic activity. The devices may be used individually, employed as sensors, used in arrays for a single specific type of particle or for a range of particle types, or configured into arrays of sensors having both these attributes.

Single particle electrochemical sensors and methods of utilization

An electrokinetic pump capable of producing high pressure is combined with a nozzle having a submicron orifice to provide a high pressure spray device. Because of its small size, the device can be contained within medical devices such as an endoscope for delivering biological materials such as DNA, chemo therapeutic agents, or vaccines to tissues and cells.

Electrokinetically pumped high pressure sprays

A device for interrupting or throttling undesired ionic transport through a fluid network is disclosed. The device acts as a fluid valve by reversibly generating a fixed 'bubble' in the conducting solvent solution carried by the network. The device comprises a porous hydrophobic structure filling a portion of a connecting channel within the network and optionally incorporates flow restrictor elements at either end of the porous structure that function as pressure isolation barriers, and a fluid reservoir connected to the region of the channel containing the porous structure. Also included is a pressure pump connected to the fluid reservoir. The device operates by causing the pump to vary the hydraulic pressure to a quantity of solvent solution held within the reservoir and porous structure. At high pressures, most or all of the pores of the structure are filled with conducting liquid so the ionic conductance is high. At lower pressures, only a fraction of the pores are filled with liquid, so ionic conductivity is lower. Below a threshold pressure, the porous structure contains only vapor, so there is no liquid conduction path. The device therefore effectively throttles ionic transport through the porous structure and acts as a 'conductance valve' or &#x201c;pressure-to-conductance&#x201d; transducer within the network.

Conductance valve and pressure-to-conductance transducer method and apparatus

An electrokinetic high pressure hydraulic pump for manipulating fluids in capillary-based system. The pump uses electro-osmotic flow to provide a high pressure hydraulic system, having no moving mechanical parts, for pumping and/or compressing fluids, for providing valve means and means for opening and closing valves, for controlling fluid flow rate, and manipulating fluid flow generally and in capillary-based systems (microsystems), in particular. The compact nature of the inventive high pressure hydraulic pump provides the ability to construct a micro-scale or capillary-based HPLC system that fulfills the desire for small sample quantity, low solvent consumption, improved efficiency, the ability to run samples in parallel, and field portability. Control of pressure and solvent flow rate is achieved by controlling the voltage applied to an electrokinetic pump.

Electrokinetic high pressure hydraulic system

A method for separating and concentrating charged species from uncharged or neutral species regardless of size differential. The method uses reversible electric field induced retention of charged species, that can include molecules and molecular aggregates such as dimers, polymers, multimers, colloids, micelles, and liposomes, in volumes and on surfaces of porous materials. The retained charged species are subsequently quantitatively removed from the porous material by a pressure driven flow that passes through the retention volume and is independent of direction thus, a multi-directional flow field is not required. Uncharged species pass through the system unimpeded thus effecting a complete separation of charged and uncharged species and making possible concentration factors greater than 1000-fold.

Electrokinetic concentration of charged molecules

A method for eliminating gas bubble blockage of current flow during operation of an electrokinetic pump. By making use of the ability to modify the surface charge on the porous dielectric medium used in electrokinetic pumps, it becomes possible to place electrodes away from the pressurized region of the electrokinetic pump. While gas is still generated at the electrodes they are situated such that the generated gas can escape into a larger buffer reservoir and not into the high pressure region of the pump where the gas bubbles can interrupt current flow. Various combinations of porous dielectric materials and ionic conductors can be used to create pumps that have desirable electrical, material handling, and flow attributes.

Method for eliminating gas blocking in electrokinetic pumping systems

Growing community of inventors

Oakland, CA, United States of America

Joseph S Schoeniger