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Genetic engineering

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https://www.readbyqxmd.com/read/28319856/systems-biology-solutions-for-biochemical-production-challenges
#1
REVIEW
Anne Sofie Lærke Hansen, Rebecca M Lennen, Nikolaus Sonnenschein, Markus J Herrgård
There is an urgent need to significantly accelerate the development of microbial cell factories to produce fuels and chemicals from renewable feedstocks in order to facilitate the transition to a biobased society. Methods commonly used within the field of systems biology including omics characterization, genome-scale metabolic modeling, and adaptive laboratory evolution can be readily deployed in metabolic engineering projects. However, high performance strains usually carry tens of genetic modifications and need to operate in challenging environmental conditions...
March 16, 2017: Current Opinion in Biotechnology
https://www.readbyqxmd.com/read/28319854/extremely-thermophilic-energy-metabolisms-biotechnological-prospects
#2
REVIEW
Christopher T Straub, Benjamin M Zeldes, Gerrit J Schut, Michael Ww Adams, Robert M Kelly
New strategies for metabolic engineering of extremely thermophilic microorganisms to produce bio-based fuels and chemicals could leverage pathways and physiological features resident in extreme thermophiles for improved outcomes. Furthermore, very recent advances in genetic tools for these microorganisms make it possible for them to serve as metabolic engineering hosts. Beyond providing a higher temperature alternative to mesophilic platforms, exploitation of strategic metabolic characteristics of high temperature microorganisms grants new opportunities for biotechnological products...
March 16, 2017: Current Opinion in Biotechnology
https://www.readbyqxmd.com/read/28319448/rationale-and-design-of-a-phase-1-clinical-trial-to-evaluate-hsv-g207-alone-or-with-a-single-radiation-dose-in-children-with-progressive-or-recurrent-malignant-supratentorial-brain-tumors
#3
Alicia M Waters, James M Johnston, Alyssa T Reddy, John Fiveash, Avi Madan-Swain, Kara Kachurak, Asim K Bag, G Yancey Gillespie, James M Markert, Gregory K Friedman
Primary central nervous system tumors are the most common solid neoplasm of childhood and the leading cause of cancer-related death in pediatric patients. Survival rates for children with malignant supratentorial brain tumors are poor despite aggressive treatment with combinations of surgery, radiation, and chemotherapy, and survivors often suffer from damaging lifelong sequelae from current therapies. Novel innovative treatments are greatly needed. One promising new approach is the use of a genetically engineered, conditionally replicating herpes simplex virus (HSV) that has shown tumor-specific tropism and potential efficacy in the treatment of malignant brain tumors...
March 2017: Human Gene Therapy. Clinical Development
https://www.readbyqxmd.com/read/28318334/development-of-lactococcus-lactis-encoding-fluorescent-proteins-gfp-mcherry-and-irfp-regulated-by-the-nisin-controlled-gene-expression-system
#4
E Martinez-Jaramillo, R Garza-Morales, M J Loera-Arias, O Saucedo-Cardenas, R Montes-de-Oca-Luna, L R McNally, J G Gomez-Gutierrez
Fluorescent proteins are useful reporter molecules for a variety of biological systems. We present an alternative strategy for cloning reporter genes that are regulated by the nisin-controlled gene expression (NICE) system. Lactoccocus lactis was genetically engineered to express green fluorescent protein (GFP), mCherry or near-infrared fluorescent protein (iRFP). The reporter gene sequences were optimized to be expressed by L. lactis using inducible promoter pNis within the pNZ8048 vector. Expression of constructions that carry mCherry or GFP was observed by fluorescence microscopy 2 h after induction with nisin...
March 20, 2017: Biotechnic & Histochemistry: Official Publication of the Biological Stain Commission
https://www.readbyqxmd.com/read/28317852/a-novel-biocontainment-strategy-makes-bacterial-growth-and-survival-dependent-on-phosphite
#5
Ryuichi Hirota, Kenji Abe, Zen-Ichiro Katsuura, Reiji Noguchi, Shigeaki Moribe, Kei Motomura, Takenori Ishida, Maxym Alexandrov, Hisakage Funabashi, Takeshi Ikeda, Akio Kuroda
There is a growing demand to develop biocontainment strategies that prevent unintended proliferation of genetically modified organisms in the open environment. We found that the hypophosphite (H3PO2, HPt) transporter HtxBCDE from Pseudomonas stutzeri WM88 was also capable of transporting phosphite (H3PO3, Pt) but not phosphate (H3PO4, Pi), suggesting the potential for engineering a Pt/HPt-dependent bacterial strain as a biocontainment strategy. We disrupted all Pi and organic Pi transporters in an Escherichia coli strain expressing HtxABCDE and a Pt dehydrogenase, leaving Pt/HPt uptake and oxidation as the only means to obtain Pi...
March 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28317440/potent-immune-responses-induced-by-a-salmonella-ghost-delivery-system-that-expresses-the-recombinant-stx2eb-fedf-and-feda-proteins-of-the-escherichia-coli-producing-f18-and-shiga-toxin-in-a-murine-model-and-evaluation-of-its-protective-effect-as-a-porcine-vaccine
#6
Gayeon Won, John Hwa Lee
BACKGROUND: In the pathogenicity of porcine edema disease, which is caused by the Escherichia coli-producing F18 and Shiga toxin, F18(+) fimbrial adhesins and Shiga toxin 2e (Stx2e) play pivotal roles in the colonization and enterotoxicity of this pathogen. OBJECTIVE: To develop a vaccine candidate against edema disease by combining three selected antigens of F18(+) Shiga toxin-producing Escherichia coli (STEC). METHODS: Genetically engineered Salmonella Typhimurium (ST) ghosts that express Stx2eB, FedF, and FedA were individually inserted in a ghost plasmid cassette (pJHL184), and the resultant plasmids were transformed into a Δasd Δlon ΔcpxR ST (JOL912)...
March 20, 2017: Veterinary Quarterly
https://www.readbyqxmd.com/read/28315813/the-case-for-applying-tissue-engineering-methodologies-to-instruct-human-organoid-morphogenesis
#7
REVIEW
Carlos R Marti-Figueroa, Randolph S Ashton
Three-dimensional organoids derived from human pluripotent stem cell (hPSC) derivatives have become widely used in vitro models for studying development and disease. Their ability to recapitulate facets of normal human development during in vitro morphogenesis produces tissue structures with unprecedented biomimicry. Current organoid derivation protocols primarily rely on spontaneous morphogenesis processes to occur within 3-D spherical cell aggregates with minimal to no exogenous control. This yields organoids containing microscale regions of biomimetic tissues, but at the macroscale (i...
March 15, 2017: Acta Biomaterialia
https://www.readbyqxmd.com/read/28315763/picw-orthologs-from-spruce-with-differential-freezing-tolerance-expressed-in-escherichia-coli
#8
Zhao Qihong, Liu Jie, Xu Xiao, Xu Qian, Gao Wei, Xu Jichen
Spruce can grow at an extra low temperature (LT), and was inferred with important antifreezing gene resources. The research here identified 4 different spruce varieties, named as PicW1, PicW2, PicM and PicK. Sequence alignment showed base-substitution and deficiency mutations among them with sequence identity between 97.61%-99.25%. Each gene was transferred into E. coli, where proteins induced by IPTG (isopropyl-β-d-thiogalactoside). Strains cultured at -5°C showed the lethal dose 50% (LD-50) between 53h-57h for the transgenic strains, but 35h for the control...
March 15, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28315745/production-of-low-expressing-recombinant-cationic-biopolymers-with-high-purity
#9
Xuguang Chen, Alireza Nomani, Niket Patel, Arash Hatefi
The growing complexity of recombinant biopolymers for delivery of bioactive agents requires the ability to control the biomaterial structure with high degree of precision. Genetic engineering techniques have provided this opportunity to synthesize biomaterials in an organism such as E. coli with full control over their lengths and sequences. One class of such biopolymers is recombinant cationic biopolymers with applications in gene delivery, regenerative medicine and variety of other biomedical applications...
March 15, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28315412/engineering-bacterial-translation-initiation-do-we-have-all-the-tools-we-need
#10
REVIEW
Justin Vigar, Hans-Joachim Wieden
BACKGROUND: Reliable tools that allow precise and predictable control over gene expression are critical for the success of nearly all bioengineering applications. Translation initiation is the most regulated phase during protein biosynthesis, and is therefore a promising target for exerting control over gene expression. At the translational level, the copy number of a protein can be fine-tuned by altering the interaction between the translation initiation region of an mRNA and the ribosome...
March 14, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/28314175/choosing-wisely-preclinical-test-models-in-the-era-of-precision-medicine
#11
REVIEW
Konrad Klinghammer, Wolfgang Walther, Jens Hoffmann
Through the introduction of a steadily growing variety of preclinical test models drug development and biomarker research has advanced. Next to classical used 2D cell line cultures, tissue-slice cultures, 3D organoid cell cultures, genetically engineered mouse models, cell line derived mouse models and patient derived xenografts may be selected for a specific question. All models harbor advantages and disadvantages. This review focuses on the available preclinical test models, novel developments such as humanized mice and discusses for which question a particular model should be employed...
March 6, 2017: Cancer Treatment Reviews
https://www.readbyqxmd.com/read/28303994/association-of-il-6-c-174g-rs-1800795-single-nucleotide-polymorphism-with-type-2-diabetes-mellitus-in-pakistani-population
#12
Amina Nadeem, Sadaf Mumtaz, Abdul Khaliq Naveed, Qaiser Mansoor, Muhammad Aslam, Arif Siddiqui, Muhammad Ismail
OBJECTIVE: To determine the association of interleukin-6 C-174G single nucleotide polymorphism with type 2 diabetes mellitus and metabolic parameters. METHODS: This case-control study was conducted from June 2012 to December 2013 at the Military Hospital Rawalpindi, the Centre for Research in Experimental and Applied Medicine, Army Medical College, Rawalpindi, and the Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan. Two cohorts of subjects were genotyped for the single nucleotide polymorphism...
March 2017: JPMA. the Journal of the Pakistan Medical Association
https://www.readbyqxmd.com/read/28303661/immunological-and-physiological-observations-in-baboons-with-life-supporting-genetically-engineered-pig-kidney-grafts
#13
Hayato Iwase, Hidetaka Hara, Mohamed Ezzelarab, Tao Li, Zhongqiang Zhang, Bingsi Gao, Hong Liu, Cassandra Long, Yi Wang, Amy Cassano, Edwin Klein, Carol Phelps, David Ayares, Abhinav Humar, Martin Wijkstrom, David K C Cooper
BACKGROUND: Genetically engineered pigs could provide a source of kidneys for clinical transplantation. The two longest kidney graft survivals reported to date have been 136 and 310 days, but graft survival >30 days has been unusual until recently. METHODS: Donor pigs (n=4) were on an α1,3-galactosyltransferase gene-knockout (GTKO)/human complement regulatory protein (CD46) background (GTKO/CD46). In addition, the pigs were transgenic for at least one human coagulation regulatory protein...
March 17, 2017: Xenotransplantation
https://www.readbyqxmd.com/read/28303150/development-of-synebrick-vectors-as-a-synthetic-biology-platform-for-gene-expression-in-synechococcus-elongatus-pcc-7942
#14
Wook Jin Kim, Sun-Mi Lee, Youngsoon Um, Sang Jun Sim, Han Min Woo
Cyanobacteria are oxygenic photosynthetic prokaryotes that are able to assimilate CO2 using solar energy and water. Metabolic engineering of cyanobacteria has suggested the possibility of direct CO2 conversion to value-added chemicals. However, engineering of cyanobacteria has been limited due to the lack of various genetic tools for expression and control of multiple genes to reconstruct metabolic pathways for biochemicals from CO2. Thus, we developed SyneBrick vectors as a synthetic biology platform for gene expression in Synechococcus elongatus PCC 7942 as a model cyanobacterium...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28302645/pd-l1-on-tumor-cells-is-sufficient-for-immune-evasion-in-immunogenic-tumors-and-inhibits-cd8-t-cell-cytotoxicity
#15
Vikram R Juneja, Kathleen A McGuire, Robert T Manguso, Martin W LaFleur, Natalie Collins, W Nicholas Haining, Gordon J Freeman, Arlene H Sharpe
It is unclear whether PD-L1 on tumor cells is sufficient for tumor immune evasion or simply correlates with an inflamed tumor microenvironment. We used three mouse tumor models sensitive to PD-1 blockade to evaluate the significance of PD-L1 on tumor versus nontumor cells. PD-L1 on nontumor cells is critical for inhibiting antitumor immunity in B16 melanoma and a genetically engineered melanoma. In contrast, PD-L1 on MC38 colorectal adenocarcinoma cells is sufficient to suppress antitumor immunity, as deletion of PD-L1 on highly immunogenic MC38 tumor cells allows effective antitumor immunity...
March 16, 2017: Journal of Experimental Medicine
https://www.readbyqxmd.com/read/28302588/nuclear-transformation-and-functional-gene-expression-in-the-oleaginous-microalga-monoraphidium-neglectum
#16
Daniel Jaeger, Wolfgang Hübner, Thomas Huser, Jan H Mussgnug, Olaf Kruse
Photosynthetic microalgae hold great promise as non-food feedstocks for the sustainable production of a range of bio-products and genetic engineering is an increasingly important strategy to improve the natural cellular features. For the vast majority of microalgal strains, however, genetic engineering is not yet possible and the establishment of efficient genetic tools for a broad range of microalgal species is an important task to reach biotechnological success. Stable DNA transformation is one of the crucial steps for most genetic engineering approaches...
March 13, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28302483/production-of-an-active-form-of-vitamin-d2-by-genetically-engineered-cyp105a1
#17
Kaori Yasuda, Yuya Yogo, Hiroshi Sugimoto, Hiroki Mano, Teisuke Takita, Miho Ohta, Masaki Kamakura, Shinichi Ikushiro, Kiyoshi Yasukawa, Yoshitsugu Shiro, Toshiyuki Sakaki
Our previous studies revealed that CYP105A1 can convert vitamin D3 (VD3) to its active form, 1α,25-dihydroxyvitamin D3 (1,25D3). Site-directed mutagenesis of CYP105A1 based on its crystal structure dramatically enhanced its activity; the activity of double variants R73A/R84A and R73A/R84V was more than 100-fold higher than that of the wild type of CYP105A1. In contrast, these variants had a low ability to convert vitamin D2 (VD2) to 1α,25-dihydroxyvitamin D2 (1,25D2), whereas they catalyzed the sequential hydroxylation at positions C25 and C26 to produce 25,26D2...
March 13, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28302114/implications-of-evolutionary-engineering-for-growth-and-recombinant-protein-production-in-methanol-based-growth-media-in-the-yeast-pichia-pastoris
#18
Josef W Moser, Roland Prielhofer, Samuel M Gerner, Alexandra B Graf, Iain B H Wilson, Diethard Mattanovich, Martin Dragosits
BACKGROUND: Pichia pastoris is a widely used eukaryotic expression host for recombinant protein production. Adaptive laboratory evolution (ALE) has been applied in a wide range of studies in order to improve strains for biotechnological purposes. In this context, the impact of long-term carbon source adaptation in P. pastoris has not been addressed so far. Thus, we performed a pilot experiment in order to analyze the applicability and potential benefits of ALE towards improved growth and recombinant protein production in P...
March 17, 2017: Microbial Cell Factories
https://www.readbyqxmd.com/read/28300077/translational-reprogramming-in-tumour-cells-can-generate-oncoselectivity-in-viral-therapies
#19
Eneko Villanueva, Pilar Navarro, Maria Rovira-Rigau, Annarita Sibilio, Raúl Méndez, Cristina Fillat
Systemic treatment of cancer requires tumour-selective therapies that eliminate cancer cells yet preserve healthy tissues from undesired damage. Tumoral transformation is associated with profound effects in translational reprogramming of gene expression, such that tumour-specific translational regulation presents an attractive possibility for generating oncoselective therapies. We recently discovered that mRNA translational control by cytoplasmic polyadenylation element-binding proteins (CPEBs) is reactivated in cancer...
March 16, 2017: Nature Communications
https://www.readbyqxmd.com/read/28299901/dual-display-of-proteins-on-the-yeast-cell-surface-simplifies-quantification-of-binding-interactions-and-enzymatic-bioconjugation-reactions
#20
Sungwon Lim, Jeff E Glasgow, Maria Filsinger Interrante, Erica M Storm, Jennifer R Cochran
Yeast surface display, a well-established technology for protein analysis and engineering, involves expressing a protein of interest as a genetic fusion to either the N- or C-terminus of the yeast Aga2p mating protein. Historically, yeast-displayed protein variants are flanked by peptide epitope tags that enable flow cytometric measurement of construct expression using fluorescent primary or secondary antibodies. Here, we built upon this technology to develop a new yeast display strategy that comprises fusion of two different proteins to Aga2p, one to the N-terminus and one to the C-terminus...
March 16, 2017: Biotechnology Journal
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