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THERMOSTABLE RNA-GUIDED ENDONUCLEASES AND METHODS OF USE THEREOF (GeoCas9)

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} The CRISPR-Cas system is now understood to confer bacteria and archaea with acquired immunity against phage and viruses. CRISPR-Cas systems consist of Cas proteins, which are involved in acquisition, targeting and cleavage of foreign DNA or RNA, and a CRISPR array, which includes direct repeats flanking short spacer sequences that guide Cas proteins to their targets. The programmable nature of these systems has facilitated their use as a versatile technology that is revolutionizing the field of genome manipulation. There is a need in the art for additional CRISPR-Cas systems with improved cleavage and manipulation under a variety of conditions and ones that are particularly thermostable under those conditions.     UC researchers discovered a new type of RNA-guided endonuclease (GeoCas9) and variants of GeoCas9.  GeoCas9 was found to be stable and enzymatically active in a temperature range of from 15°C to 75°C and has extended lifetime in human plasma.  With evidence that GeoCas9 maintains cleavage activity at mesophilic temperatures, the ability of GeoCas9 to edit mammalian genomes was then assessed.  The researchers found that when comparing the editing efficiency for both GeoCas9 and SpyCas9, similar editing efficiencies by both proteins were observed, demonstrating that GeoCas9 is an effective alternative to SpyCas9 for genome editing in mammalian cells.  Similar to CRISPR-Cas9, GeoCas9 enzymes are expected to have a wide variety of applications in genome editing and nucleic acid manipulation.   

Utilization Of Recombinant Glucosyltransferases For Value-Added Chemicals

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Calibri",sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} Glycoyltransferases are a large class of enzymes that act to catalyze the ligation of sugar onto an acceptor molecule – a process termed glycosylation. Within plants, a majority of these enzymes are involved in adding sugar groups to small molecules, forming “glycosides”. Such a modification can heavily impact the bioactivity, solubility, and physical properties of a molecule. Previous researchers have shown direct microbial bioconversion of aromatic/aliphatic flavor and fragrant molecules into their glucosides via glycosyltransferase activity via either feeding/bioconversion or direct production from glucose. However, very little emphasis has been placed on industrial yeast-­based production of specialist fragrances/flavorings or medicinal drugs.   Researchers at the University of California, Berkeley have developed a novel technology for producing plant pigment glucosides (such as highly decorated anthocyanins, coumarin glucosides, or betanins) in S. cerevisiae for industrial fermentation. Production of such colorimetric glycoside agents has value for various industries including solar-­cell, diagnostic reagent, and food-­dye manufacturers.  The technology can be used to improve the titers of commodity chemicals or the properties of various specialty or medicinal compounds. The technology also addresses one possible solution to combating the contamination of industrial fermenters through providing a method of enabling the utilization of broad-spectrum antimicrobial agents without harming the production host and as one facet of improving microbial tolerance to lignocellulose hydrolysate phenolics.  

Single Crystal Transition Metal Dichalcogenide Grown In “Jelly”

Growing single crystal material has long been a challenging problem and generally requires strict condition control.  The state-of-art methods to grow single crystal layers are performed at high temperature with high pressure or other extreme conditions. Growth of single crystal semiconductor is even more complicated and difficult. Developing a forgiving and tolerant method for single crystal growth brings great technical promise in the semiconductor industry. Transition metal dichalcogenides (TMDC) have become the most popular 2D materials that show direct bandgap when scaled down to single layer. Great effort has been put into developing synthesis techniques of large scale uniform single crystalline TMDC, including chemical vapor deposition, annealing from solution, and ALD, all of which are expensive and require high temperature. MoS2 as a TMDC family member has been widely studied for its unique properties of valley polarization, selective photoluminescence and high carrier mobility in broad range applications of valleytroncis, photonics and electronic devices. Developing methods to grow single crystal layer MoS2 will greatly reform the device fabrication process and promote its application in the semiconductor industry. Researchers at UC Berkeley have developed a strategy to grow single crystal semiconductor in a CVD coupled self-assembly process, especially for 2d materials with the assistance of self-assembly. They further develop the recipe to grown monolayer and few layer MoS2, with same mechanism but diluted solution and the assistance of surfactant. They have further patterned a thin film transistor on few layers of MoS2.

Sensitive Detection Of Chemical Species Using A Bacterial Display Sandwich Assay

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Calibri; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} Endocrine disrupting compounds are found in increasing amounts in our environment, originating from pesticides, plasticizers, and pharmaceuticals, among other sources. These compounds have been implicated in diseases such as obesity, diabetes, and cancer. The list of chemicals that disrupt normal hormone function is growing at an alarming rate, making it crucially important to find sources of contamination and identify new compounds that display this ability. However, there is currently no broad-spectrum, rapid test for these compounds, as they are difficult to monitor because of their high potency and chemical dissimilarity.   To address this, UC Berkeley researchers have developed a new detection system and method for the sensitive detection of trace compounds using electrochemical methods.  This platform is both fast and portable, and it requires no specialized skills to perform. This system enables both the detection of many detrimental compounds and signal amplification from impedance measurements due to the binding of bacteria to a modified electrode. The researchers were able to test the system finding sub-ppb levels of estradiol and ppm levels of bisphenol A in complex solutions. This approach should be broadly applicable to the detection of chemically diverse classes of compounds that bind to a single receptor.  

Epitaxial Ferroelectric On Flexible Substrate

Recent trends in electronics allude to a human-centric computing paradigm where high performance electronic devices will have to work on unusual surfaces with unconventional form factors. A key component of such a computer is a memory device for which Ferroelectric (FE) materials have long been considered as an ideal candidate. However, integration of the best quality FE films on flexible substrates has remained a daunting challenge, severely limiting the performance that can be achieved in these devices. Motivated by this challenge, UC Berkeley researchers have developed a pathway for integrating epitaxial quality, FE memory devices onto flexible substrates by providing an epitaxially grown ferroelectric stack on a flexible substrate that exhibits high performance characteristics such as high polarization, fast switching and low power operation for memory devices.

Voltage-Sensitive Dyes In Living Cells

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Calibri; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} Comprehensively mapping and recording the electrical inputs and outputs of multiple neurons simultaneously with cellular spatial resolution and millisecond time resolution remains an outstanding challenge in the field of neurobiology. Traditionally, electrophysiology is used to directly measure membrane potential changes. While this technique yields sensitive results, it is invasive and only permits single-cell recording.  VoltageFluor dyes rely on photoinduced electron transfer to effectively report membrane potential changes in cells. This approach allows for fast, sensitive and non-invasive recording of neuronal activity in cultured mammalian neurons and in ex-vivo tissue slices. However, one major limitation of small-molecule dye imaging is the inability to target the dye to specific cells of interest.   UC Berkeley researchers have developed latent voltage sensitive dyes that require a fluorogenic activation step. This new class of VoltageFluor dyes are only weakly fluorescent until being activated in defined cell types via biological processes. In particular, the VoltageFluor dyes described herein comprise a bioreversible group that quenches the fluorescence of the VoltageFluor dye, that upon selective removal by the action of biological processes (e.g., enzymes) thereby activates the fluorescence of the VoltageFluor dye. The researchers found that the new dye facilitated the observation of spontaneous activity in rat hippocampal neurons.  

Method For Imaging Neurotransmitters In Vitro and In Vivo Using Functionalized Carbon Nanotubes

Neurotransmitters play a central role in complex neural networks by serving as chemical units of neuronal communication.  Quantitative optical methods for the detection of changes in neurotransmitter levels has the potential to profoundly increase our understanding of how the brain works. Therapeutic drugs that target neurotransmitter release are used ubiquitously to treat a vast array of brain and behavioral disorders.  For example, new methods in this sphere could provide a new platform by which to validate the function of drugs that alter modulatory neurotransmission, or to screen antipsychotic and antidepressant drugs.  However, currently in neuroscience, few optical methods exist that can detect neurotransmitters with high spatial and temporal resolution in vitro or in vivo.  Brain tissue also readily scatters visible wavelengths of light currently used to perform biological imaging, and neuronal tissue and has an abundance of biomolecules that are chemically or structurally similar and therefore hard to specifically distinguish.  Furthermore, neurotransmission relevant processes occur at challenging spatial  and temporal scales.    UC Berkeley investigators have developed polymer-functionalized carbon nanotubes for in vitro and in vivo quantification of extracellular modulatory neurotransmitter levels using optical detectors. The method uses the fluorescent optical properties of polymer-functionalized carbon nanotubes to selectively report changes in concentration of specific neurotransmitters. The scheme is novel in that the detection method applies to wide variety of specific neurotransmitters, it is an optical method and therefore gives greater spatial information, and enables the potential for imaging of one or more neurotransmitters. The optical method also produces less damage to the surrounding tissue than methods that implant electrodes or cells and allows high resolution localization with other methods of optical investigation. The invention takes advantage of favorable fluorescence properties of carbon nanotubes, such as carbon nanotube emission in the near infrared and infinite fluorescence lifetime.  The near infrared emission scatters less than shorter wavelengths, enabling greater signal recovery from deeper tissue, and allows greater compatibility with other techniques. The optical properties also enable long term potentially even chronic use. 

Highly Stable Nanoscale Disk Assemblies Of The Tobacco Mosaic Virus For Applications In Drug Delivery And Disease Imaging

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Calibri; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} Self-assembling protein nanomaterials derived from viruses have properties that make them useful for applications in drug delivery, disease imaging and diagnostics. These properties include uniform sizes and shapes, biodegradability, and multiple sets of functional handles for chemical manipulation. Intact virus nanoparticles have been functionalized for applications in drug delivery in vivo, however, the injection of replication-competent viruses into subjects have limited their clinical appeal. The development of spherical and rod-shaped virus nanoparticles has in both cases resulted in differential tumor accumulation, demonstrating the need to further expand the shape library of protein nanomaterials. However, expressing non-spherical virus-based protein nanomaterials without the genetic material that functions as a backbone to the assembly architecture can lead to significant challenges including poly-diversity in size and shape, and change in assembly behavior in response to different conditions such as pH and ionic strength.   UC Berkeley researchers have developed a self-assembling nanoscale disk derived from a mutant of a recombinantly expressed viral coat protein. The disks display highly stable double-disk assembly states. The researchers functionalized the disks with the chemotherapy drug doxorubicin (DOX) and further modified the disks for improved solubility.  The functionalized disks displayed cytotoxic properties similar to those of DOX alone when incubated with U87MG glioblastoma cells, but the unmodified disks did not cause any cytotoxicity.

Robust And Selective Solid Catalyst For Tail End Of Olefin-Epoxidation Flow Reactor

Flow reactors are a useful method for Olefin epoxidation reactions, which are highly exothermic reactions.  Organic hydroperoxide and olefin conversion levels to epoxide are low at the entrance of the reactors and improve at the tail end of the reactor.  At the tail end of the reactor, there is excess alcohol coproduct and hydroperoxide in addition to epoxide.  Both Solid and liquid catalysts are used to improve conversion levels at all stages in the reactors.  The catalysts to date are efficient at the entrance of the reactor, but lose efficiency at the tail end of the reactor where epoxide is to be produced and separated.   Researchers at UC Berkeley have developed a crystalline solid catalyst for olefin epoxidation which is highly selective for epoxide production at the extreme conditions of high temperature and organic-hydroperoxide conversion at the tail end of the olefin-epoxidation reactor.  The catalyst is white crystalline solid of titanium and is based on a layered zeolite precursor.  The researchers have further developed methods of using multiple catalysts in a single reactor, where the developed catalyst is used as the catalyst at the tail end of the reactor, in the form of a packed bed, while one or more other catalyst(s) are used at the entrance of the reactor.

Coordinative Alignment Of Molecules In Chiral Metal Organic Frameworks

Single-crystal x-ray diffraction is a powerful technique for the definitive identification of chemical structures.  Although most molecules and molecular complexes can be crystallized, often enthalpic and entropic factors introduce orientational disorder that prevent determination of a high-resolution structure.  Several strategies based on the inclusion of guests in a host framework that helps maintain molecular orientation have been used to overcome this challenge.  However, most of these methods rely primarily on weak interactions to induce crystalline order of the included molecules. Researchers at UC Berkeley have developed a strategy for crystallization of molecules within the pores of chiral metal-organic frameworks (MOFs) using coordinative bonding, which includes covalent and ionic bonds, and/or using chirality.  

Enzymatic Synthesis Of Cyclic Dinucleotides

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Calibri; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} GGDEF domain-containing enzymes are diguanylate cyclases that produce cyclic di-GMP (cdiG), a second messenger that modulates the key bacterial lifestyle transition from a motile to sessile biofilm-forming state. The ubiquity of genes encoding GGDEF proteins in bacterial genomes has established the dominance of cdiG signaling in bacteria. A subfamily of GGDEF enzymes synthesizes the asymmetric signaling molecule cyclic AMP-GMP. Hybrid CDN-producing and promiscuous substrate-binding (Hypr) GGDEF enzymes are widely distributed and found in other deltaproteobacteria and have roles that include regulation of cAG signaling.  GGDEF enzymes that produce cyclic dinucleotides are especially of interest.    UC Berkeley researcher have developed a new method of preparing and using cyclic dinucleotides (CDNs) by contacting a CDN producing-enzyme (e.g., a GGDEF enzyme) with a precursor of a CDN under conditions sufficient to convert the precursor into a CDN. This method produces a variety of non-naturally occurring, asymmetric and symmetric CDNs and can be performed in vitro or in a genetically modified host cell. Also provided are CDN compositions that find use in a variety of applications such as modulating an immune response in an individual.  

Direct Optical Visualization Of Graphene On Transparent Substrates

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Calibri; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} The ∼10% optical contrast of graphene on specialized substrates like oxide-capped silicon substrates, together with the high-throughput and noninvasive features of optical microscopy, have greatly facilitated the use and research of graphene research for the past decade.  However, substantially lower contrast is obtained on transparent substrates. Visualization of nanoscale defects in graphene, e.g., voids, cracks, wrinkles, and multilayers, formed during either growth or subsequent transfer and fabrication steps, represents yet another level of challenge for most device substrates.     UC Berkeley researchers have developed a facile, label-free optical microscopy method to directly visualize graphene on transparent inorganic and polymer substrates at 30−40% image contrast per graphene layer.  Their noninvasive approach overcomes typical challenges associated with transparent substrates, including insulating and rough surfaces, enables unambiguous identification of local graphene layer numbers and reveals nanoscale structures and defects with outstanding contrast and throughput. We thus demonstrate in situ monitoring of nanoscale defects in graphene, including the generation of nano-cracks under uniaxial strain, at up to 4× video rate.  

Printable Repulsive-Force Electrostatic Actuator Methods and Device

Flexible electrostatic actuators are well designed for a range of commercial applications, from small micro-mechanical robotics to large vector displays or sound wall systems. Electrostatic actuation provides efficient, low-power, fast-response driving and control of movable nano-, micro-, and macro-structures. While commercially available electrostatic actuators have the requisite high levels of mechanical energy / force for some applications, their energy requirements are typically orders of magnitude higher than what is needed in large-area, low-power applications. Moreover, conventional approaches to these types of electrostatic actuators have limited design geometries and are prone to reliability issues like electrical shorts. To address these problems, researchers at the University of California, Berkeley, have experimented with planar electrostatic actuators using novel printing and electrode patterning and engineering techniques. The team has demonstrated a repulsive-force electrostatic actuator device (100 mm x 60 mm achieved) with extremely high field strength and high voltage operation and without insulator coatings or air breakdown.

Cas13a/C2c2 - A Dual Function Programmable RNA Endoribonuclease

Bacterial adaptive immune systems employ CRISPRs and CRISPR-associated (Cas) proteins for RNA-guided nucleic acid cleavage. Although generally targeted to DNA substrates, the Type VI CRISPR system directs interference complexes against single-stranded RNA substrates and in Type VI CRISPR systems, the single-subunit Cas13a/C2c2 protein functions as an RNA-guided RNA endonuclease.   UC Berkeley researchers have discovered that the CRISPR-Cas13a/C2c2 has two distinct RNase activities that enable both single stranded target RNA detection and multiplexed guide-RNA processing.  These dual RNase functions were found to be chemically and mechanistically different from each other and from the CRISPR RNA processing behavior of the evolutionarily unrelated CRISPR enzyme Cpf1.  Methods for detecting the single stranded target RNA were also discovered using a Cas13a/C2c2 guide RNA and a Cas13a/C2c2 protein in a sample have a plurality of RNAs as well as methods of cleaving a precursor Cas13a/C2c2 guide RNA into two or more Cas13a/C2c2 guide RNAs.  

Methods and Compositions for Increasing Desiccation Tolerance In a Cell

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Calibri; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} The impact of desiccation on microorganisms such as yeasts, bacteria, and plants are extremely important in a variety of industries ranging from the food and beverage industry that rely heavily on yeast and agricultural crops.  Microorganisms can survive for a certain period of time when water is limited, but may not be able to survive severe environmental conditions when desiccation tolerance is low. The market potential in stabilization of cells and cell products is estimated to be some $500 billion worldwide. For example, it has been reported that fewer than one in a million yeast cells from low-density logarithmic cultures of Saccharomyces cerevisiae survive desiccation. Therefore, given the exceedingly large number of microorganisms used in a variety of industries, even minor increases in survival can result in significant improvements in final output. For example, applications such as freeze-drying cells for the medical industry are used to preserve cell structure and function for long term storage. Additionally, the largest market for freeze-drying is the food industry.   UC Berkeley researchers have developed methods and compositions for increasing desiccation tolerance in a cell by contacting the cell with one or more agents that generates synergistic amounts of trehalose and a hydrophilin protein within the cell.  Cells with increased desiccation tolerance have also been developed.  

Improved Synthesis of Linear Alpha Olefins from Ethylene

Ethylene is widely used in the chemical industry for synthesis of a variety important chemicals and materials. Current ethylene-derived products include poly(ethylene), ethylene glycol, vinyls, and styrene, among others. Linear alpha olefins (LAOs) are used as comonomers in the production of linear low density poly(ethylene) (LLDPE), synthetic lubricants, and plasticizer alcohols. Current industrial processes require significant amounts of energy to produce LAOs from ethylene. Another problem relates to the generation of lower-value poly(ethylene) as insoluble solids in the reactor. To address these problems, researchers at the University of California, Berkeley, have developed a new highly selective catalytic process for synthesizing 1-hexene from ethylene. The team has demonstrated a metal-based process using a novel series of ligands, which are easily assembled from commercially available starting materials. Given the data collected to date, this inventions shows promise towards developing efficient catalytic processes for transforming ethylene into C6 or C8 LAOs.

Low-variability, Self-assembled Monolayer Doping Methods

Semiconductor materials are fundamental materials in all modern electronic devices. Continuous demand for faster and more energy-efficient electronics is pushing miniaturization and scaling to unprecedented levels. Controlled and uniform doping of semiconductor materials with atomic accuracy is critical to materials and device performance. In particular, junction depth and dopant concentration need to be tightly controlled to minimize contact resistance, as well as variability effects due to random dopant fluctuations in the channel. Conventional doping methods such as ion implantation is imprecise and can have large variability effect. Moreover, energetic introduction of dopant species will often cause crystal damage, leading to incompatibility with nanostructured-materials and further performance degradation. To address these problems, researchers at the University of California, Berkeley, have experimented with an alternative approach to a wafer-scale surface doping technique first developed at the UC Berkeley in 2007. The team has demonstrated a controlled approach for monolayer doping (MLD) in which gas phase dopant-containing molecules form low-variability, self-assembled monolayers (SAM) on target semiconductor surfaces.

PHOTO-INDUCED ELECTRON TRANSFER VOLTAGE SENSITIVE DYES

The development of fluorescent indicators for sensing membrane potential can be a challenge.  Traditional methods to measure membrane potential rely on invasive electrodes, however, voltage imaging with fluorescent probes (VF) is an attractive solution because voltage imaging circumvents problems of low- throughput, low spatial resolution, and high invasiveness. Previously reported VF probes/dyes have proven useful in a number of imaging contexts. However, the design scheme for VF dyes remains elusive, due in part to our incomplete understanding of the biophysical properties influencing voltage sensitivity in our VoltageFluor scaffolds.   UC Berkeley researchers have discovered new VF dyes, which are a small molecule platform for voltage imaging that operates via a photoinduced electron transfer (PeT) quenching mechanism to directly image transmembrane voltage changes.   The dyes further our understanding of the roles that membrane voltage plays, not only in excitable cells, such as neurons and cardiomyocytes, but also in non-excitable cells in the rest of the body.

Chemical Cocktail For Deriving Myogenic Cells

In postnatal life, growth and repair of skeletal muscle fibers are mediated by the satellite cells. These cells divide at a slow rate to sustain both self-renewal and growth of skeletal muscle tissue. In response to muscle injury, satellite cells divide and fuse to repair or replace the damaged muscular fibers. However, the self-renewal potential of adult satellite cells is limited and is compromised with aging, excessive trauma, or genetic defect as in certain severe muscular dystrophies such as Duchenne muscular dystrophy. In such cases, external interventions are needed.             UC Berkeley researchers have developed a chemical cocktail that allows large number of myogenic stem cells to be derived from, but no limited to, mouse dermal fibroblasts. These myogenic stem cells could then be transplanted into diseased or injured skeletal muscle to promote regeneration and recovery. In addition, the chemicals could be directly delivered into diseased or injured skeletal muscle to promote regeneration in vivo.  The mixture allows large number of patient-specific skeletal muscle cells to be obtained conveniently from non-invasive skin biopsy techniques. The in vitro culture of these skeletal muscle cells can then be used for disease modeling and drug screening purposes.

ENGINEERED MICROORGANISMS FOR PRODUCTION OF COMMODITY CHEMICALS AND CELLULAR BIOMASS

96 Normal 0 false false false EN-US X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Calibri; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;} Citrus pulp and sugar beet pulp are pectin-rich agricultural wastes that are globally produced in significant amounts and have the potential to contribute towards the greater bioeconomy as a source of raw, inexpensive carbohydrate biomass. There is currently limited use for these waste streams. In some cases, pulps are dried, pelleted, and repurposed as an inexpensive livestock feed, however this application is barely profitable due to high production costs. There is a need for technologies that can cost-effectively transform pectin-rich waste streams into value-added products of commercial interest.   UC Berkeley researchers developed an efficient microbial strain technology and metabolic fermentation methods for the bioconversion of pectin-rich waste streams to useful bio-based commodity chemicals and biofuels. In addition to the beneficial environmental impact of utilizing a waste-stream, the fermentation technologies achieve three design goals set to optimize the productivity of bioconversions and economic viability. First, the technology allows for anaerobic fermentation, eliminating the need for culture oxygenation. This lowers operating costs by simplifying the metabolic requirements of high-density fermentation cultures. Second, co- utilization of the major component monosaccharides in the hydrolysate broth allows for productive conversion of the predominant, energy- rich biomass sugars. Third, fermentations can be conducted at low pH, discouraging contaminant growth and eliminating the need to buffer the hydrolysate mixture.  

Metal-Organic Frameworks for H2 Adsorption and Drug Delivery

Metal–organic frameworks (MOFs) are an important class of materials with high internal surface areas and tunable pore environments that make them of interest for a wide variety of potential applications, including gas adsorption and drug delivery. One of the most ubiquitous MOF materials is of the type M2(dobdc) (2,5-dioxido-1,4-benzenedicarboxylate), sometimes referred to as M-MOF-74. The pores of these frameworks can be expanded while preserving the parent framework structure by using ligands and other analogues with multiple phenylene groups.   With an interest in exploring new ligands for expanded MOF-74 architectures, UC Berkeley researschers created a new family of expanded MOF-74 materials using the anti-inflammatory olsalazine acid as a ligand to form M2(olz), where M = Mg, Fe, Co, Ni, and Zn. Upon activation, these materials exhibit the highest Langmuir surface areas among bioactive frameworks. The M2(olz) frameworks contain pore apertures of approximately 27 Å, corresponding to the mesoporous range (≥20 Å). Strong H2 adsorption was observed by gas adsorption studies and in situ infrared spectroscopy, confirming the presence of open metal sites for all but the Zn analogue. The Mg2(olz) framework, which disassembles under physiological conditions to release olsalazine, represents an unprecedented level of loading in a bioactive metal–organic framework of 86 wt % drug. In addition to delivery of olsalazine, the large pores of Mg2(olz) were used to encapsulate a second drug, illustrating the potential of this platform to deliver multiple therapeutic components.  

Atom Probe Tomography Method and Algorithm

Most cluster analysis parameters in atom probe tomography (APT) are selected ad hoc. This can often lead to data misinterpretation and misleading results by instrument technicians and researchers. Moreover, arbitrary cluster parameters can have suboptimal consequences on data quality and integrity, leading to inefficiencies for downstream data users. To address these problems, researchers at the University of California, Berkeley, have developed a framework and specific cluster analysis methods to efficiently extract knowledge from better APT data. By using parameter selection protocols with theoretical explanations, this technology allows for a more optimized and robust multivariate statistical analysis technique from the start, thus improving the quality of analysis and outcomes for both upstream and downstream data users.

Universal Coating Compound

Polydimethyl siloxane (PDMS) has many characteristics that make it the most popular candidate for producing organ-on-a-chip devices or mirco-physiological systems (MPS) devices. After crosslinking, PDMS has shown to be biologically compatible and amenable to many standard cell culture techniques due to it’s transparency, oxygen permeability, and low auto-fluorescence. However, due to PDMS’s hydrophobicity, molecules that are also hydrophobic partition into the PDMS to produce unpredictable concentrations in cell and media channels making it impossible to predict the actual dosing concentrations for drug investigations. This unpredictability is an obstacle for using organ-on-a-chip devices as screens for drug candidates in discovery stages.   Researchers at UC Berkeley have developed a simple coating procedure that allows the formation of substrate independent (universal) coatings. The researchers identified a novel compound able to form stable coatings that outperformed existing dip-coating precursor molecules in their ability to prevent absorbance of small molecules into a variety of organic and inorganic polymers, such as PDMS. 

CRISPR genome editing of Zygotes (CRISPR-EZ)

0 0 1 214 1224 UC Berkeley 10 2 1436 14.0 Normal 0 false false false EN-US JA X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:"Table Normal"; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-parent:""; mso-padding-alt:0in 5.4pt 0in 5.4pt; mso-para-margin:0in; mso-para-margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:Cambria; mso-ascii-font-family:Cambria; mso-ascii-theme-font:minor-latin; mso-hansi-font-family:Cambria; mso-hansi-theme-font:minor-latin;} Easily accessible and efficient methodologies to edit the genomes of organisms are an immense resource to the biological and biomedical research community. Traditionally, engineering of the mammalian genome is achieved by homologous recombination (HR)-mediated sequence substitution in embryonic stem cells (ESCs), a time consuming process that occurs at low frequency. Taking genetically engineering in mice for example, after extensive screening for ESC colonies with the desired genetic modifications, ESCs are microinjected into mouse blastocysts to generate chimeras capable of germline transmission. Such chimera mice are then crossed to wild-type mice to generate heterozygous offspring (F1), which are then intercrossed to yield homozygous mutant mice (F2) that can be subjected to phenotypic analyses. Despite the wide use of this technology to generate transgenic mice, the low efficiency of HR in ESCs, the laborious process of screening, the technical difficulty of microinjection, and the nature of the mouse life cycle make this approach a lengthy and costly process.   UC Berkeley researchers developed methods for modifying the genome of a mammalian zygote by introducing a ribonucleoprotein complex (RNP) to the zygote via electroporation.  Suitable genome editing nucleases were found to be CRISPR/Cas endonucleases (e.g., class 2 CRISPR/Cas endonucleases such as a type II, type V, or type VI CRISPR/Cas endonucleases.  

Nanostructured Metal Oxide Sensing Film From Liquid Precursor

Nanostructured metal oxide materials have generated much interest for sensing applications due to their high surface area, low thermal mass, and superior performance.  However, stable and reproducible integration of these materials into a functional sensor is difficult. Vacuum deposition techniques such as sputtering or evaporation do not offer substantial sensing performance improvement. Sacrificial templating steps have been suggested, but the manufacturing complexity and cost are not suitable for high volume production. There remains a need for a simple, effective method to prepare nanostructured metal oxide films for low power, miniaturized gas sensors with high sensitivity.   Researchers at UC Berkeley have developed a novel method for creating highly porous, nanostructured metal oxide film in a controlled location from a liquid precursor using a localized heat source. This method eliminates processing steps, such as the need to separately synthesize nanomaterials and suspend them into a stable ink for deposition. The localized heat source acts to both evaporate the solvent and thermally decompose the precursor into a highly porous film of nanocrystalline metal oxide, as well as to define the location of the formed film. The utility of this method has been demonstrated for the formation of a tin oxide gas sensor with superior performance, including high sensitivity and fast response and recovery time for carbon monoxide gas. However, the method could be useful for other applications that require localized formation of a porous film of nanocrystalline metal oxide.   

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