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Synthetic transcriptional factor

Anssi Rantasalo, Christopher P Landowski, Joosu Kuivanen, Annakarin Korppoo, Lauri Reuter, Outi Koivistoinen, Mari Valkonen, Merja Penttilä, Jussi Jäntti, Dominik Mojzita
Biotechnological production of fuels, chemicals and proteins is dependent on efficient production systems, typically genetically engineered microorganisms. New genome editing methods are making it increasingly easy to introduce new genes and functionalities in a broad range of organisms. However, engineering of all these organisms is hampered by the lack of suitable gene expression tools. Here, we describe a synthetic expression system (SES) that is functional in a broad spectrum of fungal species without the need for host-dependent optimization...
June 19, 2018: Nucleic Acids Research
Smaranda Buduru, Alina-Andreea Zimta, Cristina Ciocan, Cornelia Braicu, Diana Dudea, Alexandra Iulia Irimie, Ioana Berindan-Neagoe
Over the last few decades, the incidence of oral cancer has gradually increased, due to the negative influence of environmental factors and also abnormalities within the genome. The main issues in oral cancer treatment consist in surpassing resistance and recurrence. However, continuous discovery of altered signaling pathways in these tumors provides valuable information for the identification of novel gene candidates targeted in personalized therapy. RNA interference (RNAi) is a natural mechanism that involves small interfering RNA (siRNA); this can be exploited in biomedical research by using natural or synthetic constructs for activation of the mechanism...
2018: International Journal of Nanomedicine
Daniel Bojar, Leo Scheller, Ghislaine Charpin-El Hamri, Mingqi Xie, Martin Fussenegger
Programming cellular behavior using trigger-inducible gene switches is integral to synthetic biology. Although significant progress has been achieved in trigger-induced transgene expression, side-effect-free remote control of transgenes continues to challenge cell-based therapies. Here, utilizing a caffeine-binding single-domain antibody we establish a caffeine-inducible protein dimerization system, enabling synthetic transcription factors and cell-surface receptors that enable transgene expression in response to physiologically relevant concentrations of caffeine generated by routine intake of beverages such as tea and coffee...
June 19, 2018: Nature Communications
Rachel Brough, Aditi Gulati, Syed Haider, Rahul Kumar, James Campbell, Erik Knudsen, Stephen J Pettitt, Colm J Ryan, Christopher J Lord
Although defects in the RB1 tumour suppressor are one of the more common driver alterations found in triple-negative breast cancer (TNBC), therapeutic approaches that exploit this have not been identified. By integrating molecular profiling data with data from multiple genetic perturbation screens, we identified candidate synthetic lethal (SL) interactions associated with RB1 defects in TNBC. We refined this analysis by identifying the highly penetrant effects, reasoning that these would be more robust in the face of molecular heterogeneity and would represent more promising therapeutic targets...
June 18, 2018: Oncogene
Bright N Okine, Clare Spicer, Paul Millns, Andrew Bennett, Victoria Chapman
Background and aims The clinical management of chronic neuropathic pain remains a global health challenge. Current treatments are either ineffective, or associated with unwanted side-effects. The development of effective, safe therapies requires the identification of novel therapeutic targets using clinically relevant animal models of neuropathic pain. Peroxisome proliferator activated receptor alpha (PPARα), is a member of the nuclear hormone family of transcription factors, which is widely distributed in the peripheral and central nervous systems...
December 29, 2017: Scandinavian Journal of Pain
Jurek Failmezger, Steffen Scholz, Bastian Blombach, Martin Siemann-Herzberg
Vibrio natriegens constitutes one of the fastest-growing nonpathogenic bacteria and a potential novel workhorse for many biotechnological applications. Here, we report the development of a Vibrio -based cell-free protein synthesis system (CFPS). Specifically, up to 0.4 g L-1 eGFP could be successfully synthesized in small-scale batch reactions using cell-free extract obtained from fast-growing V. natriegens cultures. Versatile CFPS system characterization attained by combining the analyses of key metabolites for translation and ribosomes revealed limitations regarding rRNA stability and critical substrate consumption (e...
2018: Frontiers in Microbiology
Alexander C Bertalovitz, Marika L Osterbur Badhey, Thomas V McDonald
Synonymous nucleotide variation is increasingly recognized as a factor than can affect protein expression, but the underlying mechanisms are incompletely understood. Here, we investigated whether synonymous changes could affect expression of the potassium voltage gated channel subfamily H member 2 (KCNH2) gene, encoding the human ether-a-go-go related gene (hERG) ion channel which is linked to hereditary cardiac arrhythmia. We examined a previously described synthetic version (hERG-CM) with synonymous substitutions designed to reduce GC content, rare codons, and mRNA secondary structure relative to the native construct (hERG-NT)...
June 15, 2018: Journal of Biological Chemistry
Laximan Sawant, Insun Kook, Jodi L Vogel, Thomas M Kristie, Clinton Jones
Following productive infection, Bovine herpesvirus-1 (BoHV-1) establishes latency in sensory neurons. Like other α-herpesviruses, expression of BoHV-1 Immediate Early (IE) genes are regulated by an enhancer complex containing the viral IE activator VP16, the cellular transcription factor Oct-1, and transcriptional coactivator HCF-1 that is assembled on an IE enhancer core element (TAATGARAT). Expression of the IE transcription unit that encodes the viral IE activators bICP0 and bICP4 may also be induced by the activated glucocorticoid receptor via two Glucocorticoid Response Elements (GREs) located upstream of the enhancer core...
June 13, 2018: Journal of Virology
Eiichi Shoguchi, Girish Beedessee, Ipputa Tada, Kanako Hisata, Takeshi Kawashima, Takeshi Takeuchi, Nana Arakaki, Manabu Fujie, Ryo Koyanagi, Michael C Roy, Masanobu Kawachi, Michio Hidaka, Noriyuki Satoh, Chuya Shinzato
BACKGROUND: The marine dinoflagellate, Symbiodinium, is a well-known photosynthetic partner for coral and other diverse, non-photosynthetic hosts in subtropical and tropical shallows, where it comprises an essential component of marine ecosystems. Using molecular phylogenetics, the genus Symbiodinium has been classified into nine major clades, A-I, and one of the reported differences among phenotypes is their capacity to synthesize mycosporine-like amino acids (MAAs), which absorb UV radiation...
June 14, 2018: BMC Genomics
Seaim Lwin Aye, Kei Fujiwara, Nobuhide Doi
Engineering and design of genetic circuit in living cell is critical in accessing the beneficial application of synthetic biology. Directed evolution can avoid the complicated rational design of such circuit by screening or selecting functional circuit from non-functional one. Here we proposed a positive-negative selection system for selecting a transcription factor that activates gene expression in response to arsenic in solution. First, we developed a whole cell biosensor for sensing arsenite in liquid using a regulator (ArsR) and a reporter (GFP), and evaluated its performance...
June 8, 2018: Journal of Biochemistry
Rui Wu, Lina Duan, Jose Pruneda-Paz, Dong-Ha Oh, Michael P Pound, Steve A Kay, Jose R Dinneny
The water stress-associated hormone abscisic acid (ABA) acts through a well-defined signal transduction cascade to mediate downstream transcriptional events important for acclimation to stress. Although ABA signaling is known to function in specific tissues to regulate root growth, little is understood regarding the spatial pattern of ABA-mediated transcriptional regulation. Here we describe the construction and evaluation of an ABSCISIC ACID RESPONSIVE ELEMENT (ABRE)-based synthetic promoter reporter that reveals the transcriptional response of tissues to different levels of exogenous ABA and stresses...
June 8, 2018: Plant Physiology
Shuobo Boboila, Gonzalo Lopez, Jiyang Yu, Debarshi Banerjee, Angela Kadenhe-Chiweshe, Eileen P Connolly, Jessica J Kandel, Presha Rajbhandari, Jose M Silva, Andrea Califano, Darrell J Yamashiro
Despite the identification of MYCN amplification as an adverse prognostic marker in neuroblastoma, MYCN inhibitors have yet to be developed. Here, by integrating evidence from a whole-genome shRNA library screen and the computational inference of master regulator proteins, we identify transcription factor activating protein 4 (TFAP4) as a critical effector of MYCN amplification in neuroblastoma, providing a novel synthetic lethal target. We demonstrate that TFAP4 is a direct target of MYCN in neuroblastoma cells, and that its expression and activity strongly negatively correlate with neuroblastoma patient survival...
June 7, 2018: Oncogene
Lin Zhu, Jiansheng Guo, Zhongyou Ma, Jianfei Wang, Cheng Zhou
Induction of ethylene biosynthesis by aphids increases the susceptibility of several plant species to aphids. Recent studies have indicated that some MYB transcription factors regulate the phloem-based defense against aphid infestation by modulating ethylene (ET) signaling. Arabidopsis MYB102 has previously been shown to be induced by wound signaling and regulate defense response against chewing insects. However, it remains unclear whether Arabidopsis MYB102 takes part in the defense response of plants to aphids...
June 7, 2018: Biomolecules
Shui-Xiu Li, Hao-Tian Wu, Yu-Ting Liu, Yi-Ying Jiang, Yi-Shan Zhang, Wei-Da Liu, Kun-Ju Zhu, Dong-Mei Li, Hong Zhang
Previous work has explored link between mitochondrial biology and fungal pathogenicity in F1 Fo -ATP synthase in Candida albicans . In this work we have detailed the more specific roles of the F1 Fo -ATP synthase β subunit, a key protein subunit of F1 Fo -ATP synthase. The ability to assimilate alternative carbons in glucose-limited host niches is known to be a critical factor for infection caused by opportunistic pathogens including C. albicans . The function of the F1 Fo -ATP synthase β subunit was characterized through the construction of an ATP2 gene null mutant ( atp2 Δ/Δ) and the gene-reconstituted strain ( atp2 Δ/ ATP2 ) in order to understand the link between carbon metabolism and C...
2018: Frontiers in Microbiology
V Raj, S Ojha, F C Howarth, P D Belur, S B Subramanya
The water-soluble vitamin, thiamine forms an important part of the diet because of its role in the energy metabolism. The protective effects of thiamine against diabetic vascular complications have been well documented. However, slower absorption and reduced bioavailability is a major limiting factor for its clinical use. To overcome this issue, lipid-soluble derivatives of thiamine (allithiamines) was developed. Among the many synthetic lipophilic derivatives of thiamine, benfotiamine (BFT) is regarded as the first choice based on its safety and clinical efficacy data...
May 2018: European Review for Medical and Pharmacological Sciences
Mohammad Foad Abazari, Fatemeh Soleimanifar, Maryam Nouri Aleagha, Sepehr Torabinejad, Navid Nasiri, Gholamreza Khamisipour, Javad Amini Mahabadi, Hossein Mahboudi, Seyed Ehsan Enderami, Ehsan Saburi, Javad Hashemi, Mousa Kehtari
Pancreatic differentiation of stem cells will aid treatment of patients with type I diabetes mellitus (T1DM). Synthetic biopolymers utilization provided extracellular matrix (ECM) and desired attributes in vitro to enhance conditions for stem cells proliferation, attachment and differentiation. A mixture of polycaprolactone and polyvinyl alcohol (PCL/PVA)-based scaffold, could establish an in vitro three-dimensional (3D) culture model. The objective of this study was investigation of the human induced pluripotent stem cells (hiPSCs) differentiation capacity to insulin-producing cells (IPCs) in 3D culture were compared with conventional culture (2D) groups evaluated at the mRNA and protein levels by quantitative PCR and immunofluorescence assay, respectively...
May 31, 2018: Gene
Kimiko Ohgi, Hiroshi Kajiya, Kazuko Goto-T, Fujio Okamoto, Yasunori Yoshinaga, Koji Okabe, Ryuji Sakagami
BACKGROUND: Lectin-like oxidized low-density-lipoprotein receptor 1 (Lox-1) is the receptor for oxidized low-density lipoprotein (oxLDL), a mediator in dyslipidemia. Toll-like receptor (TLR)-2 and - 4 are receptors of lipopolysaccharide (LPS) from Porphyromonas gingivalis, a major pathogen of chronic periodontitis. Although some reports have demonstrated that periodontitis has an adverse effect on dyslipidemia, little is clear that the mechanism is explained the effects of dyslipidemia on osteoclastogenesis...
June 2, 2018: Lipids in Health and Disease
Ankita Shrestha, Ahamed Khan, Nrisingha Dey
Co-ordinated transcriptional control employing synthetic promoters and transcription factors (TFs) can be used to achieve customized gene expression regulation in planta. Synthetic promoter technology has yielded a series of promoters with modified cis-regulatory elements that provide useful tools for efficient modulation of gene expression. In addition, the use of zinc fingers (ZFs), transcription activator-like effectors (TALEs), and catalytically inactive clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (dCas9) has made it feasible to engineer TFs that can produce targeted gene expression regulation; these approaches are particularly effective when artificial TFs are coupled with transcriptional activators or repressors...
May 30, 2018: Molecular Plant
Anna-Elisa Roser, Lucas Caldi Gomes, Rashi Halder, Gaurav Jain, Fabian Maass, Lars Tönges, Lars Tatenhorst, Mathias Bähr, André Fischer, Paul Lingor
Parkinson's disease (PD) is the second-most-frequent neurodegenerative disorder worldwide. One major hallmark of PD is the degeneration of dopaminergic (DA) neurons in the substantia nigra. Glial cell line-derived neurotrophic factor (GDNF) potently increases DA neuron survival in models of PD; however, the underlying mechanisms are incompletely understood. MicroRNAs (miRNAs) are small, non-coding RNAs that are important for post-transcriptional regulation of gene expression. Using small RNA sequencing, we show that GDNF specifically increases the expression of miR-182-5p and miR-183-5p in primary midbrain neurons (PMNs)...
June 1, 2018: Molecular Therapy. Nucleic Acids
Hong Wang, Rui Guo, Zhonghua Du, Ling Bai, Lingyu Li, Jiuwei Cui, Wei Li, Andrew R Hoffman, Ji-Fan Hu
The CRISPR-associated Cas9 system can modulate disease-causing alleles both in vivo and ex vivo, raising the possibility of therapeutic genome editing. In addition to gene targeting, epigenetic modulation by the catalytically inactive dCas9 may also be a potential form of cancer therapy. Granulin (GRN), a potent pluripotent mitogen and growth factor that promotes cancer progression by maintaining self-renewal of hepatic stem cancer cells, is upregulated in hepatoma tissues and is associated with decreased tumor survival in patients with hepatoma...
June 1, 2018: Molecular Therapy. Nucleic Acids
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