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Charged Membranes Incorporated With Porous Polymer Frameworks

Ion-exchange membranes have been established for a variety of industrial applications, including energy and environmental technologies related to water treatment, fuel cells, and flow batteries. However, the limited tunability and adverse ion permeability-selectivity tradeoff exhibited by traditional ion-exchange membranes limit their development. To address this limitation, researchers at UC Berkeley developed a new class of composite ion-exchange membrane materials incorporated with highly tunable porous aromatic frameworks (PAFs). The Berkeley researchers show that an assortment of PAF variants can be easily embedded into charged membranes, where the choice of PAF filler can be used to optimize the physical, ion transport, and adsorptive properties of the membrane according to their targeted application. Material characterizations indicate that numerous charged membranes embedded with PAFs exhibit excellent dispersibility, interfacial compatibility, structural flexibility, and pH stability. Proton conductivity and water uptake measurements also indicate that the exceptionally high porosity of PAFs enhances ion diffusion in membranes, while abundant, favorable PAF-polymer interactions decrease non-selective swelling pathways typically observed in highly charged ion-exchange membranes. Furthermore, adsorption experiments demonstrate that ion-selective PAFs can be embedded into charged membranes to tune the ion selectivity of the membrane and also enable their use as membrane adsorbents. Test show promise for technology to improve the general performance and tunability of ion-exchange membrane technologies.

Multifunctional Separations Using Adsorbent-Based Membranes

The selective separation of trace components of interest from various mixtures (e.g., micropollutants from groundwater, lithium or uranium from seawater, carbon dioxide from air) presents an especially pressing technological challenge. Established materials and separation processes seldom meet the performance standards needed to efficiently isolate these trace species for proper disposal or re-use. To address this issue, researchers at UC Berkeley developed a novel separation strategy in which highly selective and tunable adsorbents or adsorption sites are embedded into membranes. In this approach, the minor target species are selectively captured by the embedded adsorbents or adsorption sites while the species transport through the membrane. Simultaneously, the mixture can be purified through traditional membrane separation mechanisms. As a proof-of-concept, the researchers incorporated Hg2+-selective adsorbents into electrodialysis membranes that can simultaneously capture Hg2+ via an adsorption mechanism while desalinating water through an electrodialysis mechanism. Adsorption studies demonstrated that the embedded adsorbents maintain rapid, selective, regenerable, and high-capacity Hg2+ binding capabilities within the membrane matrix. Furthermore, when inserted into an electrodialysis setup, the composite membranes successfully capture all Hg2+ from various Hg2+-spiked water sources while permeating all other competing cations to simultaneously enable desalination. Finally, using an array of other ion-selective adsorbents, the Berkeley team showed that this strategy can in principle be applied generally to any target ion present in any water source. This multifunctional separation strategy can be applied to existing membrane processes to efficiently capture targeted species of interest, without the need for additional expensive equipment or processes such as fixed-bed adsorption columns.

Low Molecular Weight Xanthene-Based Fluorophore For Near- And Shortwave Infrared

Fluorophores are considered as optical “paints” to allow direct visualization of biological structures and processes with high spatial-temporal resolution. Small molecule fluorophores have been extensively studied and the color palette of these paints has expanded to the whole visible region of electromagnetic spectrum. Although powerful, the use of these imaging reagents is restricted to inherently transparent organisms and shallow imaging depths.   UC Berkeley researchers have discovered a low molecular weight xanthene-based fluorophore that exhibits excitation and emission profiles in the short wave near infrared region of the electromagnetic spectrum (SWIR). The fluorophore, KeTMR, was found to have an excitation and emission maxima at 853 nm and 1014 nm, respectively, Additional studies found the fluorophore had a Stokes shift of 161 nm, which is much larger than most rhodamine dyes.   

Design Random Heteropolymer To Transport Proton Selectively And Rapidly

Despite decades of effort, it remains challenging, if not impossible, to achieve similar transport performance similar to natural channels. Inspired by the known crystal structures of transmembrane channel proteins, protein sequence-structure-transport relationships have been applied to guide material design. However, producing both molecularly defined channel sizes and channel lumen surfaces that are chemically diverse and spatially heterogeneous have been out of reach. We show that a 4-monomer-based random heteropolymer (RHP) exhibits selective proton transport at a rate similar to those of natural proton channels. Statistical control over the monomer distribution in the RHP leads to well-modulated segmental heterogeneity in hydrophobicity, which facilitates the single RHP chains to insert into lipid bilayers. This in turn produces rapid and selective proton transport, despite the sequence variability among RHP chains. We have demonstrated the importance of:the adaptability enabled by the statistical similaritythe modularity afforded by monomer chemical diversity to achieve uniform behavior in heterogeneous systems. 

Single Conjugative Vector for Genome Editing by RNA-guided Transposition

The inventors have constructed conjugative plasmids for intra- and inter-species delivery and expression of RNA-guided CRISPR-Cas transposases for organism- and site-specific genome editing by targeted transposon insertion. This invention enables integration of large, customizable DNA segments (encoded within a transposon) into prokaryotic genomes at specific locations and with low rates of off-target integration.

Compact Ion Gun for Ion Trap Surface Treatment in Quantum Information Processing Architectures

Electromagnetic noise from surfaces is one of the limiting factors for the performance of solid state and trapped ion quantum information processing architectures. This noise introduces gate errors and reduces the coherence time of the systems. Accordingly, there is great commercial interest in reducing the electromagnetic noise generated at the surface of these systems.Surface treatment using ion bombardment has shown to reduce electromagnetic surface noise by two orders of magnitude. In this procedure ions usually from noble gasses are accelerated towards the surface with energies of 300eV to 2keV. Until recently, commercial ion guns have been repurposed for surface cleaning. While these guns can supply the ion flux and energy required to prepare the surface with the desired quality, they are bulky and limit the laser access, making them incompatible with the requirements for ion trap quantum computing.To address this limitation, UC Berkeley researchers have developed an ion gun that enables in-situ surface treatment without sacrificing high optical access, enabling in situ use with a quantum information processor.

Reticulation Of Macromolecules Into Crystalline Networks

Covalent organic frameworks (COFs) are 2D or 3D extended periodic networks assembled from symmetric, shape persistent molecular 5 building blocks through strong, directional bonds. Traditional COF growth strategies heavily rely on reversible condensation reactions that guide the reticulation toward a desired thermodynamic equilibrium structure. The requirement for dynamic error correction, however, limits the choice of building blocks and thus the associated mechanical and electronic properties imbued within the periodic lattice of the COF.   UC Berkeley researchers have demonstrated the growth of crystalline 2D COFs from a polydisperse macromolecule derived from single-layer graphene, bottom-up synthesized quasi one-dimensional (1D) graphene nanoribbons (GNRs). X-ray scattering and transmission electron microscopy revealed that 2D sheets of GNR-COFs self-assembled at a liquid-l quid interface stack parallel to the layer boundary and exhibit an orthotropic crystal packing. Liquid-phase exfoliation of multilayer GNR-COF crystals gave access to large area bilayer and trilayer cGNR-COF films. The functional integration of extended 1D materials into crystalline COFs greatly expands the structural complexity and the scope of mechanical and physical materials properties.

Temporal Control over DNA-Patterned Signaling Ligands In Vitro Using Sequence-Targeting Nucleases

UC Berkeley researchers have created a new technique that can rapidly “print” two-dimensional arrays of cells and proteins that mimic a wide variety of cellular environments in the body, be it the brain tissue surrounding a neural stem cell, the lining of the intestine or liver or the cellular configuration inside a tumor.  In the new technique, each cell or protein is tethered to a substrate with a short string of DNA. While similar methods have been developed that attach tethered cells or proteins one by one.  By repeating the process, up to 10 different kinds of cells or proteins can be tethered to the surface in an arbitrary pattern. This technique could help scientists develop a better understanding of the complex cell-to-cell messaging that dictates a cell’s final fate, from neural stem cell differentiating into a brain cell to a tumor cell with the potential to metastasize to an embryonic stem cell becoming an organ cell.

Cellular Potassium Imaging Using A Ratiometric Fluorescent Sensor

The inventors developed a ratiometric fluorescent small molecule probe for potassium ion detection composed of a duo-fluorophore system (KR-1). UV-vis detector and fluorometer measurement support ratiometric response of the probe towards potassium ion concentration. The probe was further applied to cellular potassium level detection using confocal microscope imaging technique. KR-1 enables simple determination of potassium levels in various cancer or non-cancer cell lines.

Covalent Organic Framework With Exceptional Water Sorption Properties

A new covalent organic framework (COF) with defective square lattice topology and exceptional water sorption properties stemming fro its unique framework structure. The COF exhibits a working capacity of 0.23 g(H2O)/g(COF) between 20 and 40% relative humidity without displaying hysteretic behavior. Furthermore, it maintains these promising water sorption properties after several uptake and release cycles. This material could be used as a sorbent for water harvesting or other water sorption related applications.

Low Band Gap Graphene Nanoribbon Electronic Devices

This invention creates a new graphene nanoribbons (GNR)-based transistor technology capable of pushing past currently projected limits in the operation of digital electronics for combining high current (i.e. high speed) with low-power and high on/off ratio. The inventors describe the design and synthesis of molecular precursors for low band gap armchair graphene nanoribbons (AGNRs) featuring a width of N=11 and N=15 carbon atoms, their growth into AGNRs, and their integration into functional electronic devices (e.g. transistors). N is the number of carbon atoms counted in a chain across the width and perpendicular to the long axis of the ribbon.

Illumination Device for Dynamic Spatiotemporal Control of Photostimulation

A programmable LED device that illuminates multiple spatial locations (termed wells) with user-defined light patterns whose intensity can be modulated as a function of space and time. The devices are used for optogenetic stimulation of tissue culture plates (24-well and 96-well) kept in a heated and humidified tissue culture incubator, as well as photopatterning of hydrogels. In brief, light from LEDs passes through optical elements that ensure uniform illumination of each well. Parameters of the optical system, such as LED configuration, optical diffuser elements, materials, and geometry, were modeled and optimized using the optical ray tracing software Zemax OpticStudio. An electronics subsystem allows programmed control of illumination intensity and temporal sequences, with independent control of each well. Spatial precision is conveyed through a photomask attached to the culture plate. The hardware design also includes a cooling system and vibration isolation to reduce heating and damage to the sample. Lastly, a graphical user interface (GUI) was used to wirelessly program the illumination intensity and temporal sequences for each well. The devices can thus illuminate 24 independent channels with visible, NIR, or UV light with intensity ranges of 0 to 20-100 microwatts per millimeter-squared with 16-bit intensity resolution, and a temporal resolution of 1 millisecond and spatial resolution of 100 microns. In summary, the device allows uniform illumination of multiple wells for multiplexed photoactivation or photopolymerization of various substrates (light-responsive bacterial or mammalian cells grown in tissue culture, hydrogels, dyes, etc) with user-defined patterns. The device can be combined with a robotic handler, microscope, spectrometer, etc, to enable high-throughput illumination and simultaneous recording of the sample.

Targeted Ionophore-Based Metal Supplementation

Metal deficiency is implicated in a variety of genetic, neurological, cardiovascular, and metabolic diseases. Current approaches for addressing metal deficiency rely on generic metal ion supplementation, which can potentially lead to detrimental off-target metal accumulation in unwanted tissues and subsequently trigger oxidative stress and damage cascades. The inventors have developed a new modular platform for delivering metal ions in a tissue-specific manner and demonstrate liver-targeted copper supplementation as a proof of concept of this strategy. Specifically, the inventors designed and synthesized a N-acetylgalactosamine-functionalized ionophore, Gal-Cu(gtsm), to serve as a copper-carrying “Trojan Horse” that targets liver-localized asialoglycoprotein receptors (ASGPRs) and releases copper only after being taken up by cells, where the reducing intracellular environment triggers copper release from the ionophore. The inventors utilized a combination of bioluminescence imaging and inductively-coupled plasma mass spectrometry assays to establish ASGPR-dependent copper accumulation with this reagent in both liver cell culture and mouse models with minimal toxicity. The modular nature of this synthetic approach presages that this platform can be expanded to deliver a broader range of metals to specific cells, tissues, and organs in a more directed manner to treat metal deficiency in disease. This patent broadly covers directed metal delivery to select organs, tissues, and organelles.

A Protein Inhibitor Of Cas9

  Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas9 nucleases, when complexed with a guide RNA, effect genome editing in a sequence-specific manner. RNA-guided Cas9 has proven to be a versatile tool for genome engineering in multiple cell types and organisms.  There is a need in the art for additional compositions and methods for controlling genome editing activity of CRISPR/Cas9.   UC Berkeley researchers have discovered a new protein that is able to inhibit the Cas9 protein from Staphyloccocus aureus (SauCas9). SauCas9 is smaller than the frequently used Cas9 from Streptococcus pyogenes, which has a number of benefits for delivery. The inhibitor is a small protein from a phage and is capable of strongly inhibiting gene editing in human cells.


Several chemical, physical, and biological techniques have been used for delivering macromolecules into living cells. Delivery of biomolecules into living cells is essential for biomedical research and drug development as well as genome editing. However, conventional methods of delivery of biomolecules such as viral vectors, cell penetrating peptides, cationic lipids, positive charged polymers, bulk electroporation, and microinjection pose several challenges. Such challenges include safety concerns, toxicity, damage to the cells, limited loading capacity, low delivery efficiencies, low cell viabilities, low cell throughput, high cellular perturbation, and high costs.  Thus, there is a need for delivery devices and methods that allow for permeabilization of the cell membrane to facilitate delivery of biomolecules into cells.   UC Berkeley researchers have developed a universal delivery electroporation system that makes cell transfection very simple for all of types of cells. The technology can be used to replace conventional cellular delivery methods such as cationic lipid, positive charged polymer and bulk electroporation as well as microinjection.  The system can deliver biomolecules (e.g., DNA, RNA, proteins, nucleic acid-protein complexes (e.g., RNPs)) or other reagents into all cell types, including T-cells, which cannot be efficiently transfected with conventional approaches.  

Cas12-mediated DNA Detection Reporter Molecules

Class 2 CRISPR-Cas systems are streamlined versions in which a single Cas protein (an effector protein, e.g., a type V Cas effector protein such as Cpf1) bound to RNA is responsible for binding to and cleavage of a targeted sequence. The programmable nature of these minimal systems has facilitated their use as a versatile technology that continues to revolutionize the field of genome manipulation.    Cas12 is an RNA-guided protein that binds and cuts any matching DNA sequence. Binding of the Cas12-CRISPR RNA (crRNA) complex to a matching single-stranded DNA (ssDNA) or double-stranded DNA (dsDNA) molecule activates the protein to non-specifically degrade any ssDNA in trans. Cas12a-dependent target binding can be coupled to a reporter molecule to provide a direct readout for DNA detection within a sample.  UC Berkeley researchers have developed compositions, systems, and kits having labeled single stranded reporter DNA molecules that provide a sensitive readout for detection of a target DNA. 

Combination Of Air Lubrication And Super Hydrophobic Frictional Drag Reduction

This technology combines air layer frictional drag reduction (ALDR) with super hydrophobic surfaces (SHS) to achieve frictional drag reduction of ALDR with significantly reduced gas flux. Thus, enabling increased net energy savings. The stable air layer is achieved with lesser gas flux when utilizing a SHS.Periodic air layers may replenish SHS, enabling drag reduction with reduced energy cost. Combinations of SHS and regular or other non-SHS surface may be used to control spreading of gas, thus facilitating formation of ALDR using discrete gas injection points better than previously achievable. Such surface variations could also be used to preferentially guide gas towards or away from propulsion, depending on desired outcome. By controlling ALDR regionally or globally on a surface, with or without SHS, this technology modifies flow around a hull. This mediates forces on partially or fully submerged objects, enabling control of flow patterns, resistance, steering, and/or dynamics.

Enhanced Speed Polymerases For Sanger Sequencing

Sanger sequencing has remained a dominant DNA sequencing methodology for molecular biology research and development for many years.  The main commercially available DNA polymerase developed for Sanger sequencing has a slow extension speed and has difficulties sequencing secondary structures such as GC rich regions, hairpins, mono- and poly-nucleotide repeats.  While specialized plastics and reductions in reaction volumes to improve Sanger sequencing reaction times, any gains in sequencing assay performance (e.g., sequencing time or throughput) are offset by increased costs associated with a terminator reagent.  During the last two decades, further refinement and advancement of suitable DNA polymerases to improve polymerization speeds during Sanger sequencing have been limited.  Thus, there remains a need for improved DNA polymerases suitable for Sanger sequencing that possess enhanced elongation speeds, and the ability to sequence through secondary structures present in DNA templates.    A UC Berkeley researchers has discovered compositions and methods for preparing and using Taq DNA polymerases with improved Sanger sequencing elongation sequencing rates as compared to commercially available Sanger sequencing reagents.  

Protein-Coated Microparticles For Protein Standardization In Single-Cell Assays

Single-cell analysis offers powerful capabilities of identification of rare sub-populations of cells, understanding heterogeneity of cancerous tumors, and tracking cell differentiation and reprogramming. Despite great potentials for uncovering new biological systems and targeting diseases with precision medicine, single-cell approaches are composed of complex device processes that can cause bias in measurement.  In deep sequencing, technical variation in single cell expression data occurs during capture and pre-amplification steps. Similarly, in single-cell protein assays, technical variability can obscure functionally relevant variance.    To better control protein measurement quality in single-cell assays, researchers at the University of California, Berkeley developed a novel method to loading and release protein standard. This method utilizes the surface of modified and functionalized microparticles as vehicles to capture target proteins with desired concentrations. Chelation-assisted click chemistry is applied to demonstrate that protein standards with different molecular masses can be loaded and bounded in a single-cell protein assay. Microparticles are introduced into single-cell devices by either passive gravity, magnetic attraction, or other physicochemical forces. These protein standards from microparticles provide a reference to measure protein mass sizes from individual cells and a quality control for any biases in device fabrication, cell lysis, protein solubility, protein capture, and protein readouts (i.e. antibody probing).   

Endoribonucleases For Rna Detection And Analysis

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;} Bacteria and archaea possess adaptive immune systems that rely on small RNAs for defense against invasive genetic elements. CRISPR (clustered regularly interspaced short palindromic repeats) genomic loci are transcribed as long precursor RNAs, which must be enzymatically cleaved to generate mature CRISPR-derived RNAs (crRNAs) that serve as guides for foreign nucleic acid targeting and degradation. This processing occurs within the repetitive sequence and is catalyzed by a dedicated CRISPR-associated (Cas) family member in many CRISPR systems.  Endoribonucleases that process CRISPR transcripts are bacterial or archaeal enzymes capable of catalyzing sequence- and structure- specific cleavage of a single- stranded RNA. These enzymes cleave a specific phosphodiester bond within a specific RNA sequence.  UC Berkeley researchers discovered variant Cas endoribonucleases, nucleic acids encoding the variant Cas endoribonucleases, and host cells genetically modified with the nucleic acids that can be used, potentially in conjunction with Cas9, to detect a specific sequence in a target polyribonucleotide and of regulating production of a target RNA in a eukaryotic cell.  For example, it was found that the variant Cas endoribonuclease has an amino acid substitution at a histidine residue such that is is enzymatically inactive in the absence of imidazole and is activatable in the presence of imidazole.  

Printed All-Organic Reflectance Oximeter Array

A flexible reflectance oximeter array (ROA) composed of printed organic light-emitting diodes (OLEDs) and organic photodiodes (OPDs), which senses reflected light from tissue to determine the oxygen saturation. Since reflected light is used as the signal, the sensor array can be used beyond the conventional sensing locations. We implemented the ROA to measure SpO2 on the forehead with 1.1% mean error and to create two-dimensional (2D) oxygenation maps of the adult forearm under pressure cuff-induced ischemia. Due to the mechanical flexibility, 2D oxygenation mapping capability, and the ability to place the sensor in diverse places, the ROA is promising for novel medical sensing applications such as mapping oxygenation in tissues, wounds, or transplanted organs.

Selective Nitrogen Adsorption Using a Vanadium Metal-Organic Framework

Natural gas, composed primarily of methane, has many potential uses as a cleaner and more renewable source of energy than other fossil fuels. However, about 20% of US natural gas reserves contain levels of N2 that are too high for pipeline processing. Using natural gas from renewable sources also encounters this problem. Furthermore, in processing steps to create high-purity methane from its various sources, the removal of N2 remains a significant energetic cost. This separation is typically performed through cryogenic distillation, and improvements in energy efficiency of this separation are necessary to utilize the many available sources of methane. Switching to membrane or adsorbent-based technologies could potentially alleviate this challenge. Size selective molecular sieves and membranes have demonstrated some ability for separating N2 from CH4, but face problems with scalability and selectivity; and current adsorbents need significant improvements in selectivity and capacity for N2 to be commercially viable.  To address this situation, researchers at UC Berkeley have developed a new adsorbent V2Cl2(btdd) with exceptional affinity for nitrogen, such that early experiments already demonstrate a N2/CH4 selectivity of over 10x greater than any reported material. The Berkeley material is a permanently porous vanadium(II)-containing metal-organic framework (MOF). It represents the first example of a MOF with five-coordinate vanadium(II) centers as the primary metal node. The electronic properties of these five-coordinate V(II) centers make this MOF uniquely reactive towards relatively inert and weak electron acceptors, such as nitrogen, creating a stronger M–N2 interaction than any known MOF. Additionally, the high-density of V(II) centers translates to a high gas uptake capacity, qualifying this material as a promising N2/CH4 selective adsorbant. Key performance parameters can be tuned as the building blocks are synthetically modifiable.

Type V CRISPR/CAS Effector Proteins for Cleaving ssDNA and Detecting Target DNA

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; mso-bidi-font-family:"Times New Roman"; mso-bidi-theme-font:minor-bidi;} Class 2 CRISPR–Cas systems (e.g., type V CRISPR/Cas systems such as Cas12 family systems) are characterized by effector modules that include a single effector protein. For example, in a type V CRISPR/Cas system, the effector protein - a CRISPR/Cas endonuclease (e.g., a Cas12a protein) - interacts with (binds to) a corresponding guide RNA (e.g., a Cas12a guide RNA) to form a ribonucleoprotein (RNP) complex that is targeted to a particular site in a target nucleic acid via base pairing between the guide RNA and a target sequence within the target nucleic acid molecule.  Thus, like CRISPR-Cas9, Cas12 has been harnessed for genome editing based on its ability to generate targeted, double-stranded DNA (dsDNA) breaks.   UC Berkeley researchers have discovered that RNA-guided DNA binding unleashes indiscriminate single-stranded DNA (ssDNA) cleavage activity by Cas12a that completely degrades ssDNA molecules. The researchers found that target-activated, non-specific ssDNase cleavage is also a property of other type V CRISPR-Cas12 enzymes. By combining Cas12a ssDNase activation with isothermal amplification, the researchers were able to achieve attomolar sensitivity for DNA detection.  For example, rapid and specific detection of human papillomavirus in patient samples was achieved using these methods and compositions.   

Computed Axial Lithography (CAL) For 3D Additive Manufacturing

Additive manufacturing fabrication methods are proliferating rapidly, with photopolymer-based approaches comprising some of the most prominent methods. These stereolithographic techniques provide a useful balance of resolution, build speed, process control, and capital cost (system metrics that typically must be traded off one against another). Resolving the speed limitations, surface roughness (stair-step artifacts), and requirements for support structures would provide the next major steps forward in the progress of these technologies.To address this potential, researchers at UC Berkeley have developed a system and method that accomplishes volumetric fabrication by applying computed tomography techniques in reverse, fabricating structures by exposing a photopolymer resin volume from multiple angles, updating the light field at each angle. The necessary light fields are spatially and/or temporally multiplexed, such that their summed energy dose in a target resin volume crosslinks the resin into a user-defined geometry. These light-fields may be static or dynamic and may be generated by a spatial light modulator that controls either the phase or the amplitude of a light field (or both) to provide the necessary intensity distribution.


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.   

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