Technologies

Research Tools

Most Recent Inventions

S1mplex: A New Tool for Precision Gene Editing

UW–Madison researchers have developed a modular RNA aptamer-streptavidin strategy, termed S1mplex, to ‘sharpen the scalpels’ used in genome surgery. In the new approach, CRISPR-Cas9 RNPs are complexed with a nucleic acid donor template, as well as other biotinylated molecules (e.g., quantum dots).

In human pluripotent stem cells, tailored S1mplexes increased the ratio of precisely edited to imprecisely edited alleles up to 18-fold higher than standard gene editing methods, and enriched cell populations containing multiplexed precise edits up to 42-fold.

These advances with versatile, preassembled reagents could greatly reduce the time and cost of in vitro/ex vivo gene editing applications in precision medicine and drug discovery, and aid in the development of increased and multiple dosing regimens for somatic gene editing in vivo.
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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.
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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.
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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.
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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.
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Most Recent Patents

Improved Methods and Materials for Transforming Plant Cells

UW-Madison researchers have developed a method of using a DNA-launching platform to introduce viral RNA into a host cell that has been engineered to support viral replication and expression. The platform encodes a modified viral RNA molecule located downstream of a DNA-dependent RNA polymerase promoter. After the platform is introduced into a host cell, it effectively “launches” the RNA molecule into the cell, where the RNA is replicated and expressed.
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Poly(UG) Polymerase: A Useful New RNA Tool

UW–Madison researchers have identified a poly(UG) polymerase in a roundworm called Caenorhabditis elegans. The newly discovered enzyme adds repeating UG sequences to the ends of RNA. This activity could be useful as a research tool in vitro, e.g., providing a new way to synthesize cDNA of RNAs of unknown sequence.

The gene in C. elegans that encodes the enzyme is called RDE-3. Although its sequence was already known, its polymerase activity was not.
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Bio-Based Production of Non-Straight-Chain and Oxygenated Fatty Acids for Fuels and More

UW–Madison researchers have identified several enzymes in the bacterium Rhodobacter sphaeroides that can be purified to produce non-straight-chain fatty acids in vitro or expressed in genetically modified microorganisms including E. coli for synthesis in vivo. Strains may be ‘fine-tuned’ to produce a specific type of non-straight-chain fatty acid (e.g., furan-containing) by expressing, overexpressing or deleting the enzymes in various combinations.
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