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Genetic engineering

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https://www.readbyqxmd.com/read/29145596/a-crispr-cas9-based-exploration-into-the-elusive-mechanism-for-lactate-export-in-saccharomyces-cerevisiae
#1
Robert Mans, Else-Jasmijn Hassing, Melanie Wijsman, Annabel Giezekamp, Jack T Pronk, Jean-Marc Daran, Antonius J A van Maris
CRISPR/Cas9-based genome editing allows rapid, simultaneous modification of multiple genetic loci in Saccharomyces cerevisiae. Here, this technique was used in a functional analysis study aimed at identifying the hitherto unknown mechanism of lactate export in this yeast. First, an S. cerevisiae strain was constructed with deletions in 25 genes encoding transport proteins, including the complete aqua(glycero)porin family and all known carboxylic-acid transporters. The 25-deletion strain was then transformed with an expression cassette for Lactobacillus casei lactate dehydrogenase (LcLDH)...
November 14, 2017: FEMS Yeast Research
https://www.readbyqxmd.com/read/29145364/could-genetic-engineering-save-the-gal%C3%A3-pagos
#2
Stephen S Hall
No abstract text is available yet for this article.
November 14, 2017: Scientific American
https://www.readbyqxmd.com/read/29145021/production-of-chemicals-using-dynamic-control-of-metabolic-fluxes
#3
REVIEW
Peng Xu
Engineered microbial cell factories are constantly experiencing metabolic imbalance due to nutrients depletion, metabolites buildup, evolutionary pressure or genetic instability. It is important to equip the engineered cell factory with sensor-regulator system to enable cell adjust metabolism and respond to the changing environment. Dynamically allocating cellular resources and optimally controlling pathway expression have proved as promising strategies to manage the tradeoff between cell growth and product formation as well as improve the cost-competitiveness of industrial fermentation...
November 13, 2017: Current Opinion in Biotechnology
https://www.readbyqxmd.com/read/29144959/induction-of-innervation-by-encapsulated-adipocytes-with-engineered-vitamin-a-metabolism
#4
Q Shen, R Yasmeen, J Marbourg, L Xu, J L Yu, P Fadda, A Flechtner, L J Lee, P G Popovich, O Ziouzenkova
Innervation is a fundamental basis for function and survival of tissues. In the peripheral tissues, degenerative diseases create a neurotoxic metabolic milieu that either causes neurodegeneration or fails to sustain regenerative growth and reinnervation of injured/diseased tissues. Encapsulation of cells producing neurotrophic factors can augment axon growth and neuron survival; however, sustained innervation in vivo requires a combination of factors promoting axon growth and guidance pathway that are released in a tissue-specific context...
October 28, 2017: Translational Research: the Journal of Laboratory and Clinical Medicine
https://www.readbyqxmd.com/read/29144774/strategies-for-enhanced-crop-resistance-to-insect-pests
#5
Angela E Douglas
Insect pests are responsible for substantial crop losses worldwide through direct damage and transmission of plant diseases, and novel approaches that complement or replace broad-spectrum chemical insecticides will facilitate the sustainable intensification of food production in the coming decades. Multiple strategies for improved crop resistance to insect pests, especially strategies relating to plant secondary metabolism and immunity and microbiome science, are becoming available. Recent advances in metabolic engineering of plant secondary chemistry offer the promise of specific toxicity or deterrence to insect pests; improved understanding of plant immunity against insects provides routes to optimize plant defenses against insects; and the microbiomes of insect pests can be exploited, either as a target or as a vehicle for delivery of insecticidal agents...
November 16, 2017: Annual Review of Plant Biology
https://www.readbyqxmd.com/read/29144726/physalis-mottle-virus-like-particles-as-nanocarriers-for-imaging-reagents-and-drugs
#6
Hema Masarapu, Bindi K Patel, Paul L Chariou, He Hu, Neetu M Gulati, Bradley L Carpenter, Reza A Ghiladi, Sourabh Shukla, Nicole F Steinmetz
Platform technologies based on plant virus nanoparticles (VNPs) and virus-like particles (VLPs) are attracting the attention of researchers and clinicians because the particles are biocompatible, biodegradable, noninfectious in mammals, and can readily be chemically and genetically engineered to carry imaging agents and drugs. When the Physalis mottle virus (PhMV) coat protein is expressed in Escherichia coli, the resulting VLPs are nearly identical to the viruses formed in vivo. Here, we isolated PhMV-derived VLPs from ClearColi cells and carried out external and internal surface modification with fluorophores using reactive lysine-N-hydroxysuccinimide ester and cysteine-maleimide chemistries, respectively...
November 16, 2017: Biomacromolecules
https://www.readbyqxmd.com/read/29144386/creating-a-world-class-program-through-training-and-certification
#7
Ann Turner
The animal research field has gotten more sophisticated with the use of genetically engineered animals, biohazardous agents, and advanced technologies. Trained and competent personnel are a cornerstone of any animal care and use program. Individual career success is a combination of education, experience, continuing competence, professional development, and personal commitment. Integration of training and certification into programs demonstrate institutional commitment to quality research and enhance the program by providing staff with knowledge and training to address problems and situation that arise and to perform their job in a professional and effective manner...
November 16, 2017: Behavioral Sciences
https://www.readbyqxmd.com/read/29143424/restoration-of-ribozyme-tertiary-contact-and-function-using-a-molecular-glue-for-rna
#8
Chikara Dohno, Maki Kimura, Kazuhiko Nakatani
Some RNA classes require folding into the proper higher-order structures to exert their functions. Hammerhead ribozyme (HHR) requires a folding conformation stabilized by tertiary interaction for full activity. Here, we developed a rationally engineered HHR that was inactive, but could be activated by a synthetic RNA-binding ligand, naphthyridine carbamate tetramer with Z-stilbene linker (Z-NCTS). Binding of Z-NCTS could induce the formation of an active folding structure and thereby restore ribozyme activity, where Z-NCTS acts as a molecular glue to bring two isolated RNA loops into contact with each other...
November 16, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/29143082/spatial-separation-of-photosynthesis-and-ethanol-production-by-cell-type-specific-metabolic-engineering-of-filamentous-cyanobacteria
#9
Shigeki Ehira, Takuto Takeuchi, Akiyoshi Higo
Cyanobacteria, which perform oxygenic photosynthesis, have drawn attention as hosts for the direct production of biofuels and commodity chemicals from CO2 and H2O using light energy. Although cyanobacteria capable of producing diverse chemicals have been generated by metabolic engineering, anaerobic non-photosynthetic culture conditions are often necessary for their production. In this study, we conducted cell type-specific metabolic engineering of the filamentous cyanobacterium Anabaena sp. PCC 7120, which forms a terminally differentiated cell called a heterocyst with a semi-regular spacing of 10-15 cells...
November 15, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/29142828/metabolic-engineering-of-ustilago-trichophora-tz1-for-improved-malic-acid-production
#10
Thiemo Zambanini, Hamed Hosseinpour Tehrani, Elena Geiser, Christiane K Sonntag, Joerg M Buescher, Guido Meurer, Nick Wierckx, Lars M Blank
Ustilago trichophora RK089 has been found recently as a good natural malic acid producer from glycerol. This strain has previously undergone adaptive laboratory evolution for enhanced substrate uptake rate resulting in the strain U. trichophora TZ1. Medium optimization and investigation of process parameters enabled titers and rates that are able to compete with those of organisms overexpressing major parts of the underlying metabolic pathways. Metabolic engineering can likely further increase the efficiency of malate production by this organism, provided that basic genetic tools and methods can be established for this rarely used and relatively obscure species...
June 2017: Metabolic Engineering Communications
https://www.readbyqxmd.com/read/29141886/pd-l1-genetic-overexpression-or-pharmacological-restoration-in-hematopoietic-stem-and-progenitor-cells-reverses-autoimmune-diabetes
#11
Moufida Ben Nasr, Sara Tezza, Francesca D'Addio, Chiara Mameli, Vera Usuelli, Anna Maestroni, Domenico Corradi, Silvana Belletti, Luca Albarello, Gabriella Becchi, Gian Paolo Fadini, Christian Schuetz, James Markmann, Clive Wasserfall, Leonard Zon, Gian Vincenzo Zuccotti, Paolo Fiorina
Immunologically based clinical trials performed thus far have failed to cure type 1 diabetes (T1D), in part because these approaches were nonspecific. Because the disease is driven by autoreactive CD4 T cells, which destroy β cells, transplantation of hematopoietic stem and progenitor cells (HSPCs) has been recently offered as a therapy for T1D. Our transcriptomic profiling of HSPCs revealed that these cells are deficient in programmed death ligand 1 (PD-L1), an important immune checkpoint, in the T1D nonobese diabetic (NOD) mouse model...
November 15, 2017: Science Translational Medicine
https://www.readbyqxmd.com/read/29140662/re-framing-biotechnology-regulation
#12
Alison Peck
Biotechnology is about to spill the banks of federal regulation. New genetic engineering techniques like CRISPR-Cas9 promise revolutionary breakthroughs in medicine, agriculture, and public health—but those techniques would not be regulated under the terms of the Coordinated Framework for Regulation of Biotechnology. This revolutionary moment in biotechnology offers an opportunity to correct the flaws in the framework, which was hastily patched together at the advent of the technology. The framework has never captured all relevant technologies, has never satisfied the public that risk is being effectively managed, and has never been accessible to small companies and publicly-funded labs that increasingly are positioned to make radical, life-saving innovations...
2017: Food and Drug Law Journal
https://www.readbyqxmd.com/read/29140243/a-versatile-genetic-tool-for-post-translational-control-of-gene-expression-in-drosophila-melanogaster
#13
Sachin Sethi, Jing W Wang
Several techniques have been developed to manipulate gene expression temporally in intact neural circuits. However, the applicability of current tools developed for in vivo studies in Drosophila is limited by their incompatibility with existing GAL4 lines and side effects on physiology and behavior. To circumvent these limitations, we adopted a strategy to reversibly regulate protein degradation with a small molecule by using a destabilizing domain (DD). We show that this system is effective across different tissues and developmental stages...
November 15, 2017: ELife
https://www.readbyqxmd.com/read/29138316/germline-cas9-expression-yields-highly-efficient-genome-engineering-in-a-major-worldwide-disease-vector-aedes-aegypti
#14
Ming Li, Michelle Bui, Ting Yang, Christian S Bowman, Bradley J White, Omar S Akbari
The development of CRISPR/Cas9 technologies has dramatically increased the accessibility and efficiency of genome editing in many organisms. In general, in vivo germline expression of Cas9 results in substantially higher activity than embryonic injection. However, no transgenic lines expressing Cas9 have been developed for the major mosquito disease vector Aedes aegypti Here, we describe the generation of multiple stable, transgenic Ae. aegypti strains expressing Cas9 in the germline, resulting in dramatic improvements in both the consistency and efficiency of genome modifications using CRISPR...
November 14, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29138304/refactoring-the-genetic-code-for-increased-evolvability
#15
Gur Pines, James D Winkler, Assaf Pines, Ryan T Gill
The standard genetic code is robust to mutations during transcription and translation. Point mutations are likely to be synonymous or to preserve the chemical properties of the original amino acid. Saturation mutagenesis experiments suggest that in some cases the best-performing mutant requires replacement of more than a single nucleotide within a codon. These replacements are essentially inaccessible to common error-based laboratory engineering techniques that alter a single nucleotide per mutation event, due to the extreme rarity of adjacent mutations...
November 14, 2017: MBio
https://www.readbyqxmd.com/read/29137675/human-hyaluronic-acid-synthase-1-promotes-malignant-transformation-via-epithelial-to-mesenchymal-transition-micronucleation-and-centrosome-abnormalities
#16
Nguyet Nguyen, Awanit Kumar, Simi Chacko, Rodney J Ouellette, Anirban Ghosh
BACKGROUND: Human hyaluronic acid (HA) molecules are synthesized by three membrane spanning Hyaluronic Acid Synthases (HAS1, HAS2 and HAS3). Of the three, HAS1 is found to be localized more into the cytoplasmic space where it synthesizes intracellular HA. HA is a ubiquitous glycosaminoglycan, mainly present in the extracellular matrix (ECM) and on the cell surface, but are also detected intracellularly. Accumulation of HA in cancer cells, the cancer-surrounding stroma, and ECM is generally considered an independent prognostic factors for patients...
November 14, 2017: Cell Communication and Signaling: CCS
https://www.readbyqxmd.com/read/29137648/lesr-is-a-novel-upstream-regulator-that-controls-downstream-clp-expression-to-modulate-antibiotic-hsaf-biosynthesis-and-cell-aggregation-in-lysobacter-enzymogenes-oh11
#17
Huiyong Xu, Ruping Wang, Yangyang Zhao, Zheng Qing Fu, Guoliang Qian, Fengquan Liu
BACKGROUND: Heat-stable antifungal factor (HSAF) is a polycyclic tetramate macrolactam secondary metabolite that exhibits broad-spectrum inhibitory activities against filamentous fungal pathogens. The native yield of this chemical is low. It is also a great challenge to synthesize HSAF artificially, due to its complex structure. Understanding the regulatory mechanism underlying HSAF biosynthesis could provide genetic basis for engineering high HSAF-producing strain. The transcription factor Clp is a global regulator that controls bacterial pathogenicity and the expression of one hundred related genes in the phytopathogenic bacterium Xanthomonas campestris pv...
November 14, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/29137643/efficient-gene-editing-in-corynebacterium-glutamicum-using-the-crispr-cas9-system
#18
Feng Peng, Xinyue Wang, Yang Sun, Guibin Dong, Yankun Yang, Xiuxia Liu, Zhonghu Bai
BACKGROUND: Corynebacterium glutamicum (C. glutamicum) has traditionally been used as a microbial cell factory for the industrial production of many amino acids and other industrially important commodities. C. glutamicum has recently been established as a host for recombinant protein expression; however, some intrinsic disadvantages could be improved by genetic modification. Gene editing techniques, such as deletion, insertion, or replacement, are important tools for modifying chromosomes...
November 14, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/29135456/probiotics-and-their-use-in-inflammatory-bowel-disease
#19
Muhammad Amer, Muhammad Nadeem, Saeed Ur Rashid Nazir, Maira Fakhar, Fatima Abid, Qurat-Ul Ain, Elliyah Asif
Context • Crohn's disease and ulcerative colitis result in similar gastrointestinal (GI) symptoms, including pain, diarrhea, stools with mucus or blood, and ulceration or tissue damage within the alimentary canal. Gut microbiota play a crucial role in triggering, maintaining, and exacerbating inflammatory bowel disease (IBD). Probiotics might help to rebalance the gut flora in a positive way, shifting from pro- to anti-inflammatory. Objectives • The study intended to investigate the safety and use of probiotics and the biological effects of probiotic bacteria on IBD...
November 14, 2017: Alternative Therapies in Health and Medicine
https://www.readbyqxmd.com/read/29134331/optimization-and-application-of-a-dna-launched-infectious-clone-of-equine-arteritis-virus
#20
Ting Qi, Xiaojun Wang
Reverse genetics is one of the most powerful tools in modern virology. Equine arteritis virus (EAV) is the prototype member of the Equartevirus. In this study, a new reverse genetics system for the recovery of equine arteritis virus from a cDNA plasmid, which contains viral cDNA sequence flanked by hammerhead ribozyme (HamRz) and hepatitis delta virus ribozyme (HdvRz) sequences in both terminals of the viral genome, was developed by optimization of the promoter and terminator regions. Cellular RNA polymerase II drove the transcription of the viral genome...
November 13, 2017: Applied Microbiology and Biotechnology
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