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Modulation Of Wnt5a To Treat Glaucoma

A major risk factor for glaucoma which affects over 3 million Americans and 60 million people worldwide is increased intraocular pressure (IOP), which can damage the optic nerve and cause permanent blindness without treatment. Currently, there is no cure for glaucoma. Existing eye drops or oral medications are of limited efficacy with many side effects, and surgeries often fail with scar formation and fibrosis. Schlemm’s canal (SC) is a circumferential channel located at the iridocorneal angle in the ocular anterior chamber. It is part of the conventional aqueous humor outflow system, which accounts for 70–90% of the total aqueous humor that drains out of the eye in human. The endothelial cell lining of Schlemm’s canal is one of the primary sites of resistance to aqueous humor drainage and is a major determinant of IOP. When canal resistance increases with age or under pathological situation, IOP is elevated leading to glaucoma with irreversible optic nerve damage and vision loss. It is therefore an important target for glaucoma therapy.    UC researchers have discovered that Wnt5a is expressed on Schlemm’s canal, where its expression is regulated in response to sheer stress change, and devised a method for treating Glaucoma or pathogenic intraocular pressure by locally administering to an eye in need thereof formulations of a Wnt5a inhibitor.

NANOPORE MEMBRANE DEVICE AND METHODS OF USE THEREOF

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.  

Lentivirus-like Particle Delivery of CRISPR-Cas9 & Guide RNA for Gene Editing

CRISPR-Cas9 is revolutionizing the field of gene editing and genome engineering. Efficient methods for delivering CRISPR-Cas9 genome editing components into target cells must be developed, both for ex vivo and in vivo applications. Current delivery strategies have drawbacks: genetically encoding Cas9 into viruses (ex. adeno-associated virus, adenovirus, retrovirus) leads to prolonged Cas9 expression in target cells, thus increasing the likelihood for off-target gene editing events. This problem can be mitigated by complexing ribonucleoprotein (RNP) Cas9 and guide RNA (gRNA) in vitro prior to administration – however, additional strategies for trafficking RNPs into target cells must additionally be employed.    To address this challenge, UC Berkeley researchers have discovered lentivirus-like particles that deliver Cas9/gRNA RNP complexes into target cells with high efficiency. This delivery strategy combines the ability of viruses to deliver cargo intracellularly with the transient nature of Cas9 RNP complexes. 

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.  

Integrated Nanocrescent Optical Antenna System (iNOAS) for Rapid Precision Molecular Diagnostics

UC Researchers have developed a hexagonally packed nanocrescent optical antenna array for real-time ultrafast photonic PCR, which can avoid the problem of quenching fluorescent probes by applying well-matched resonance oscillation of electrons instead of random photothermal heating. Matching the plasmon resonance with NIR diode and the design of integrated nanocrescent optical antenna on chip allows effective plasmonic conversion of the light to heat drastically. It improves not only time consuming thermal cycling step, but also real­time PCR detection due to amplified signals without the excessive heating problem of fluorescent molecules. 

Topical Anti-proliferative Agents for Melanoma

Due to the year-to-year increase in skin cancer incidences and dramatic decrease in survival, once the melanoma has metastasized, a preventative treatment for skin cancer would be significant. Currently, the only defenses against melanoma are applying sun protection factor (SPF) regularly and protecting oneself from direct sun exposure. In a 2015 national survey conducted by the Center for Disease Control and Prevention (CDC), 34% of adults reported using SPF 15 or higher and 35% of adults reported having a sunburn in the past year.    UC Berkeley researchers have discovered active antiproliferative compounds that can be applied post-sunburn to prevent the growth and metastases of melanoma and therefore, could reduce the number of melanoma incidences per year. Based on preliminary data, they are developing a compound cocktail composed of active lead compounds to develop an anti-proliferative and cytostatic topical treatment of melanoma to slow down tumor development. 

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.