A5000874602- CRISPR and Genetic Engineering
- Retinal Development and Disorders
- Neuroscience and Neural Engineering
- Pluripotent Stem Cells Research
- RNA Interference and Gene Delivery
- Genetics, Aging, and Longevity in Model Organisms
- Virus-based gene therapy research
- Cellular Mechanics and Interactions
- Photochemistry and Electron Transfer Studies
- Graphene and Nanomaterials Applications
- Proteoglycans and glycosaminoglycans research
- Advanced biosensing and bioanalysis techniques
- Lysosomal Storage Disorders Research
- Electrochemical Analysis and Applications
- Cell Adhesion Molecules Research
- Free Radicals and Antioxidants
Wisconsin Institutes for Discovery
2013-2020
University of Wisconsin–Madison
2013-2020
Smith-Kettlewell Eye Research Institute
2019-2020
Massachusetts Institute of Technology
2010
University of California, Riverside
2009
The estimation of the driving force for photoinduced charge-transfer processes, using Rehm−Weller equation, requires employment redox and spectroscopic quantities describing participating electron donor acceptor. Although data are usually obtained from diluted solutions, potentials most frequently electrochemical measurements conducted in concentrated electrolyte solutions. To correct differences media, which various types conducted, a term, based on Born equation solvation energy ions, is...
Genome-edited human pluripotent stem cells (hPSCs) have broad applications in disease modeling, drug discovery, and regenerative medicine. We present characterize a robust method for rapid, scarless introduction or correction of disease-associated variants hPSCs using CRISPR/Cas9. Utilizing non-integrated plasmid vectors that express puromycin N-acetyl-transferase (PAC) gene, whose expression translation is linked to Cas9, we transiently select based on their early levels Cas9 protein. Under...
Tissue engineering and advanced manufacturing of human stem cells requires a suite tools to control gene expression spatiotemporally in culture. Inducible systems offer cell-extrinsic control, typically through addition small molecules, but molecule inducers contain few functional groups for further chemical modification. Doxycycline (DXC), potent inducer tetracycline (Tet) transgene systems, was conjugated hyperbranched dendritic polymer (Boltorn H40) subsequently reacted with polyethylene...
CRISPR-Cas9 gene editing of human cells and tissues holds much promise to advance medicine biology, but standard methods require weeks months reagent preparation selection where or all the initial edited samples are destroyed during analysis. ArrayEdit, a simple approach utilizing surface-modified multiwell plates containing one-pot transcribed single-guide RNAs, separates thousands cell populations for automated, live, high-content imaging The lowers time cost produces embryonic stem at...
Background: Non-viral gene-editing of human cells using the CRISPR-Cas9 system requires optimized delivery multiple components. Both Cas9 endonuclease and a single guide RNA, that defines genomic target, need to be present co-localized within nucleus for efficient occur. This work describes new high-throughput screening platform optimization strategies. Methods: By exploiting high content image analysis microcontact printed plates, multi-parametric outcome data from hundreds thousands...
Abstract Dominantly inherited disorders are not typically considered therapeutic candidates for gene augmentation. Here, we utilized patient-specific induced pluripotent stem cell-derived retinal pigment epithelium (iPSC-RPE) to test the potential of augmentation treat Best disease, a dominant macular dystrophy caused by over 200 missense mutations in BEST1 . Gene iPSC-RPE fully restored calcium-activated chloride channel activity and improved rhodopsin degradation models recessive...
Gene-edited human cells are important resources for drug target identification, regulatory science, regenerative medicine and basic biology. Recently, use of the CRISPR-Cas9 system drastically cut time required to produce gene-edited cell lines down a few months. Targeted gene disruption in population followed by selection next-generation sequencing can identify targets, however many these methods destroy all mutant clones, so subsequent gene-editing experiment is obtain living downstream...
Hemodynamic forces induced by blood flow are potent regulators of vascular homeostasis and arterial structure. While previous work has focused on the role integrin receptors in mechanotransduction, little is known about cell surface proteoglycans these processes. This focuses specifically heparan sulfate proteoglycan syndecan‐1 mechanotransduction. We isolated smooth muscle cells from knockout mice tranfected them with wild‐type mutated forms using lentiviral vectors. Wild‐type,...
Dominantly inherited disorders are not typically considered therapeutic candidates for gene augmentation (GA). We tested whether GA or genome editing (GE) could serve as a solo therapy autosomal dominant Best disease (adBD), macular dystrophy linked to over 100 mutations in the BEST1 gene, which encodes homo-pentameric calcium-activated chloride channel (CaCC) retinal pigment epithelium (RPE). Since no suitable animal models of adBD exist, we generated RPE from patient-derived induced...