Research Tools

Most Recent Inventions

Designing Programmable Inducible Promoters for Biosensor Applications

UW–Madison researchers have developed a method for de novo design of synthetic inducible promoters for transcription factors and other DNA binding proteins such as aTFs with tunable dynamic range behavior and compatibility with virtually any host organism.

The method can include selecting inducible promoters, for example, by converting a constitutive promoter of an organism into an inducible promoter by introducing binding sites near the RNA polymerase binding site. By controlling the access of a transcription factor and the RNA polymerase to the promoter, the dynamic range of the system can be controlled.

Creating ‘Designer’ Yeast Hybrids for Brewing and More

UW–Madison researchers have developed HyPr, a simple and efficient method for generating synthetic Saccharomyces hybrids without sporulation or modification of the nuclear genome.

Specifically, using the new method, induction of HO endonuclease expression by a promoter in two diploid cultures, followed by co-culture and subsequent double-drug selection, will produce hybrids at a rate approaching 1 out of 1,000 cells plated. Plasmids can then be easily cured or spontaneously lost to produce strains without genome modifications.

The resulting strains can be rapidly screened for plasmid loss, opening an efficient route towards meeting the Generally Recognized as Safe (GRAS) standards of the U.S. Department of Agriculture and FDA.

Efficient In Vitro Assay for Antigen-Specific Tolerance

Building on their work, UW–Madison researchers have now developed a T cell-bound cytokine (T-CBC) assay for detecting and quantifying regulatory T cells specific to self-antigens or donor alloantigens. The new method comprises (a) culturing the subject’s T cells for 24 hours in the presence of one or more target antigens and (b) analyzing the cultured T cells for expression of a marker (EBi3; TGFβ/LAP) indicative of antigen-specific immune suppression.

Monomeric Fluorescent Protein-Ligand Complexes with Strong Fluorescence in the Far-Red Region

Research from the University of Wisconsin-Washington County in collaboration with the Institute for Stem Cell Biology and Regenerative Medicine in India, has resulted in the development of monomeric variants of the naturally occurring Sandercyanin Fluorescent Protein (SFP) using site-directed mutagenesis. This work has stemmed from earlier research focused on development of the tetrameric form of SFP, a biliverdin-binding lipocalin protein originally isolated from the mucus of the blue walleye fish, Sander vitreus. Monomeric variants of SFP (mSFPs) have been found to possess the same non-covalent, bili-binding characteristics of the tetramer but are one-quarter the size (~18.6kDa) and do not oligomerize. They are therefore anticipated to be more useful in a host of biotechnology applications. Like the tetrameric form, the mSFPs have a large stokes shift (375nm/675nm) and fluoresce in the far-red or near infrared region, which is advantageous for a wide range of applications including investigation of protein-protein interactions, spatial and temporal gene expression, assessing cell biology distribution and mobility, studying protein activity and protein interactions in vivo, as well as cancer research, immunology, and stem cell research and sub-cellular localization. In addition, the newly developed mSFP’s far-red fluorescence is particularly advantageous for in vivo, deep-tissue imaging.

Enhanced Endotoxin Detection: New Advantages in Liquid Crystal Assays for Gram-Negative Pathogens

UW–Madison researchers have now demonstrated enhanced endotoxin detection in the presence of masking agents in their previous liquid crystal system.

Unlike the LAL assay, the LC-based method does not suffer from LER or any loss of sensitivity due to the presence of cations (e.g., Ca2+ or Mg2+), buffers (e.g., citrate), surfactants (e.g., SDS), chelating agents (e.g., EDTA), proteins or nucleic acids (e.g., DNA or RNA). Thus, the LC-based method provides faster and cheaper detection of endotoxin when compared to existing methods, such as the LAL assay.

Most Recent Patents

Zip-Lignin™ Assay: An Analysis and Validation Tool

The researchers have now developed the most sensitive assay to date for detecting and quantifying Zip-lignin monomers in plants. They modified an existing lignin assay known as DFRC (Derivatization Followed by Reductive Cleavage) that has been in use for almost a decade. They incorporated several new features to improve the sensitivity of the assay, including extended incubation periods and an additional purification step.

The modified DFRC assay is currently the only known technique capable of determining levels of monolignol ester conjugates in plant lignin.

Enhanced Biomass Digestion with Wood Wasp Bacteria

UW–Madison researchers have derived preparations from ActE secretions that highly degrade lignocellulose. The bacteria can be obtained from Sirex noctilio wasps and grown on a substrate containing mostly cellulose, hemicelluloses, xylan, wood or non-wood biomass, and chitin. The substrate may be pretreated for better results. The ActE are grown aerobically to maximize the secretion of both oxidative and hydrolytic enzymes capable of rapid deconstruction of matter. The secretions can be purified and added directly to biomass slurry.

Invertebrate Models for Studying Traumatic Brain Injury

UW–Madison researchers have developed a method for inflicting closed head TBI in invertebrates. The method can be used to screen candidate therapeutic agents for treating TBI.

Specifically, the researchers created a device that mechanically induces TBI in organisms such as Drosophila melanogaster (fruit flies), which respond to impact trauma in many of the same ways as humans. Subjecting these organisms to a controlled impact and dosing with test agents could help identify the biological pathways associated with the consequences of TBI as well as enable screens for therapeutic compounds.

The device consists of a spring attached to a vial containing up to 100 flies. The spring is pulled back and then released, striking the vial against a rubber pad to inflict brain trauma as the flies collide with the bottom and walls of the vial. The device can be adjusted to inflict injury at differing severities.