A5054251605- Cancer-related Molecular Pathways
- Ubiquitin and proteasome pathways
- CRISPR and Genetic Engineering
- RNA Interference and Gene Delivery
- Genetics, Aging, and Longevity in Model Organisms
- Environmental Toxicology and Ecotoxicology
- Cancer Research and Treatments
- Advanced biosensing and bioanalysis techniques
- interferon and immune responses
- RNA and protein synthesis mechanisms
- Genetically Modified Organisms Research
- Toxic Organic Pollutants Impact
- Marine Biology and Environmental Chemistry
- Nanoparticles: synthesis and applications
- Effects and risks of endocrine disrupting chemicals
- Molecular Biology Techniques and Applications
- Cellular Mechanics and Interactions
- Marine Bivalve and Aquaculture Studies
- Algal biology and biofuel production
- Immunotherapy and Immune Responses
- Animal Genetics and Reproduction
- Cervical Cancer and HPV Research
- Plant Disease Resistance and Genetics
- RNA regulation and disease
- Mesenchymal stem cell research
University of Wyoming
2015-2023
Horizon Discovery Group (United States)
2017
Thermo Fisher Scientific (Japan)
2012
University of Maine
2000-2011
Thermo Fisher Scientific (United States)
2009
The University of Texas at Austin
2004-2008
St. Jude Children's Research Hospital
2004
Ubiquitin-(Ub) like proteins (Ubls) are conjugated to their targets by an enzymatic cascade involving E1 activating enzyme, E2 conjugating and in some cases E3 ligase. ISG15 is a Ubl that cellular after IFN-α/β stimulation. Although the enzyme for (Ube1L/E1 ) has been identified, identities of downstream components conjugation have remained elusive. Here we report purification demonstrate it UbcH8, also functions Ub conjugation. In vitro assays with purified enzymes vivo RNA interference...
During development, biomechanical forces contour the body and provide shape to internal organs. Using genetic molecular approaches in combination with a FRET-based tension sensor, we characterized pulling force exerted by elongating pharynx (foregut) on anterior epidermis during C. elegans embryogenesis. Resistance of this actomyosin-based circumferential constricting is mediated FBN-1, ZP domain protein related vertebrate fibrillins. fbn-1 was required specifically within FBN-1 expressed...
Since its initial application in mammalian cells, CRISPR-Cas9 has rapidly become a preferred method for genome engineering experiments. The Cas9 nuclease is targeted to genomic DNA using guide RNAs (gRNA), either as the native dual RNA system consisting of DNA-targeting CRISPR (crRNA) and trans-activating crRNA (tracrRNA), or chimeric single (sgRNA). Entirely DNA-free systems protein mRNA chemically synthesized gRNAs allow transient expression components, thereby reducing potential...
Most human cancer cells display increased telomerase activity that appears to be critical for continued cell proliferation and tumor formation. The E6 protein of malignancy-associated papillomaviruses increases cellular in primary keratinocytes at least partly via transcriptional activation the catalytic subunit, hTERT. In present study, we investigated whether E6AP, a ubiquitin ligase well known binding mediating some activities oncoprotein, participated transactivation hTERT promoter. Our...
The function of the human papillomavirus (HPV) E6 protein that is most clearly linked to carcinogenesis targeted degradation p53, which dependent on E6AP ubiquitin ligase. Additional functions have been attributed E6, including stimulation telomerase activity and other cellular proteins, but in cases it unclear whether these activities are also dependent. While influences transcriptional program HPV-positive cell lines through inactivation has shown at least a subset its p53-independent...
E1 and E2 enzymes coordinate the first steps in conjugation of ubiquitin (Ub) ubiquitin-like proteins (Ubls). ISG15 is an interferon-alpha/beta-induced Ubl, for are Ube1L UbcH8, respectively. UbcH7 most closely related to yet it does not function vivo, while both UbcH8 have been reported Ub conjugation. Kinetic analyses wild-type chimeric E2s were performed determine basis preferential activation by whether activated equally well E1(Ub) (Ube1). K(m) determinations confirmed strong preference...
While the use of RNA interference (RNAi) in molecular biology and functional genomics is a well-established technology, vivo applications synthetic short interfering RNAs (siRNAs) require chemical modifications. We recently found that amides as non-ionic replacements for phosphodiesters may be useful modifications optimization siRNAs. Herein, we report comprehensive study systematic replacement single phosphate with an amide linkage throughout guide strand The results show are surprisingly...
Potential in vivo applications of RNA interference (RNAi) require suppression various off-target activities. Herein, we report that replacement a single phosphate linkage between the first and second nucleosides passenger strand with an amide almost completely abolished its undesired activity restored desired guide strands had been compromised by unfavorable modifications. Molecular modeling suggested observed effect was most likely due to suppressed loading amide-modified into Ago2 caused...
The CRISPR-Cas9 system has been utilized for large-scale, loss-of-function screens mainly using lentiviral pooled formats and cell-survival phenotypic assays. Screening in an arrayed format expands the types of readouts that can be used to now include high-content, morphology-based assays, with recent availability synthetic crRNA libraries, new studies are emerging. Here, we use a cell cycle reporter line perform arrayed, crRNA:tracrRNA screen targeting 169 genes (>600 crRNAs) high content...
The Direct PCR approach facilitates amplification directly from small amounts of unpurified samples, and is demonstrated here for several plant animal tissues (Figure 1). based on specially engineered Thermo Scientific Phusion Phire DNA Polymerases, which include a double-stranded binding domain that gives them unique properties such as high tolerance inhibitors. PCR-based target detection has numerous applications in research, including genotype analysis verification transgenes....
During embryogenesis the nascent Caenorhabditis elegans epidermis secretes an apical extracellular matrix (aECM) that serves as external stabilizer, preventing deformation of by mechanical forces exerted during morphogenesis. At present, factors contribute to aECM function are mostly unknown, including components themselves, their posttranslational regulators, and pathways required for secretion. Here we showed two proteins previously linked function, SYM-3/FAM102A SYM-4/WDR44, colocalize...
The Direct PCR approach facilitates amplification directly from small amounts of unpurified samples, and is demonstrated here for several plant animal tissues (Figure 1). based on specially engineered Thermo Scientific Phusion Phire DNA Polymerases, which include a double-stranded binding domain that gives them unique properties such as high tolerance inhibitors. PCR-based target detection has numerous applications in research, including genotype analysis verification transgenes....
During embryogenesis the nascent Caenorhabditis elegans epidermis secretes an apical extracellular matrix (aECM) that serves as external stabilizer, preventing deformation of by mechanical forces exerted during morphogenesis. We showed two conserved proteins linked to this process, SYM-3/FAM102A and SYM-4/WDR44, colocalize intracellular membrane-associated puncta likely function together in a complex. Proteomics data also suggested potential roles for FAM102A WDR44 family trafficking,...