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recombinant DNA technology

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https://www.readbyqxmd.com/read/29352253/tracking-the-evolution-of-transiently-transfected-individual-cells-in-a-microfluidic-platform
#1
Micaela Tamara Vitor, Sébastien Sart, Antoine Barizien, Lucimara Gaziola De La Torre, Charles N Baroud
Transient gene expression (TGE) technology enables the rapid production of large amount of recombinant proteins, without the need of fastidious screening of the producing cells required for stable transfection (ST). However, several barriers must be overcome before reaching the production yields using ST. For optimizing the production yields from suspended cells using TGE, a better understanding of the transfection conditions at the single cell level are required. In this study, a universal droplet microfluidic platform was used to assess the heterogeneities of CHO-S population transiently transfected with cationic liposomes (CL) (lipoplexes) complexed with GFP-coding plasmid DNA (pDNA)...
January 19, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29330710/glycogen-production-in-marine-cyanobacterial-strain-synechococcus-sp-nkbg-15041c
#2
Amr Badary, Shouhei Takamatsu, Mitsuharu Nakajima, Stefano Ferri, Peter Lindblad, Koji Sode
An important feature offered by marine cyanobacterial strains over freshwater strains is the capacity to grow in seawater, replacing the need for often-limited freshwater. However, there are only limited numbers of marine cyanobacteria that are available for genetic manipulation and bioprocess applications. The marine unicellular cyanobacteria Synechococcus sp. strain NKBG 15041c (NKBG15041c) has been extensively studied. Recombinant DNA technologies are available for this strain, and its genomic information has been elucidated...
January 12, 2018: Marine Biotechnology
https://www.readbyqxmd.com/read/29318202/fundamental-crispr-cas9-tools-and-current-applications-in-microbial-systems
#3
REVIEW
Pingfang Tian, Jia Wang, Xiaolin Shen, Justin Forrest Rey, Qipeng Yuan, Yajun Yan
Derived from the bacterial adaptive immune system, CRISPR technology has revolutionized conventional genetic engineering methods and unprecedentedly facilitated strain engineering. In this review, we outline the fundamental CRISPR tools that have been employed for strain optimization. These tools include CRISPR editing, CRISPR interference, CRISPR activation and protein imaging. To further characterize the CRISPR technology, we present current applications of these tools in microbial systems, including model- and non-model industrial microorganisms...
September 2017: Synthetic and Systems Biotechnology
https://www.readbyqxmd.com/read/29316791/serine-integrases-advancing-synthetic-biology
#4
Christine Merrick, Jia Zhao, Susan Rosser
Serine integrases catalyze precise rearrangement of DNA through site-specific recombination of small sequences of DNA called attachment (att) sites. Unlike other site-specific recombinases, the recombination reaction driven by serine integrases is highly directional and can only be reversed in the presence of an accessory protein called a recombination directionality factor (RDF). The ability to control reaction directionality has led to the development of serine integrases as tools for controlled rearrangement and modification of DNA in synthetic biology, gene therapy, and biotechnology...
January 9, 2018: ACS Synthetic Biology
https://www.readbyqxmd.com/read/29250325/modern-biotechnology-based-therapeutic-approaches-against-hiv-infection
#5
Muhammad Imran, Yasir Waheed, Ayesha Ghazal, Sajjad Ullah, Sher Zaman Safi, Muhsin Jamal, Muhammad Ali, Muhammad Atif, Muhammad Imran, Farman Ullah
The causative agent of acquired immune deficiency syndrome (AIDS) is human immunodeficiency virus (HIV). Since its discovery before 30 years, a number of drugs known as highly active antiretroviral therapy have been developed to suppress the life cycle of the virus at different stages. With the current therapeutic approaches, ending AIDS means providing treatment to 35 million individuals living with HIV for the rest of their lives or until a cure is developed. Additionally, therapy is associated with various other challenges such as potential of drug resistance, toxicity and presence of latent viral reservoir...
December 2017: Biomedical Reports
https://www.readbyqxmd.com/read/29249046/generation-of-transgenic-goats-by-pronuclear-microinjection-a-retrospective-analysis-of-a-commercial-operation-1995-2012
#6
W Gavin, S Blash, N Buzzell, D Pollock, L Chen, N Hawkins, J Howe, K Miner, J Pollock, C Porter, M Schofield, Y Echelard, H Meade
Production of transgenic founder goats involves introducing and stably integrating an engineered piece of DNA into the genome of the animal. At LFB USA, the ultimate use of these transgenic goats is for the production of recombinant human protein therapeutics in the milk of these dairy animals. The transgene or construct typically links a milk protein specific promoter sequence, the coding sequence for the gene of interest, and the necessary downstream regulatory sequences thereby directing expression of the recombinant protein in the milk during the lactation period...
December 16, 2017: Transgenic Research
https://www.readbyqxmd.com/read/29247730/collapse-of-dna-in-packaging-and-cellular-transport
#7
REVIEW
T J Thomas, Thresia Thomas
The dawn of molecular biology and recombinant DNA technology arose from our ability to manipulate DNA, including the process of collapse of long extended DNA molecules into nanoparticles of approximately 100 nm diameter. This condensation process is important for the packaging of DNA in the cell and for transporting DNA through the cell membrane for gene therapy. Multivalent cations, such as natural polyamines (spermidine and spermine), were initially recognized for their ability to provoke DNA condensation...
December 13, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/29240926/exocet-exonuclease-in-vitro-assembly-combined-with-recet-recombination-for-highly-efficient-direct-dna-cloning-from-complex-genomes
#8
Hailong Wang, Zhen Li, Ruonan Jia, Jia Yin, Aiying Li, Liqiu Xia, Yulong Yin, Rolf Müller, Jun Fu, A Francis Stewart, Youming Zhang
The exponentially increasing volumes of DNA sequence data highlight the need for new DNA cloning methods to explore the new information. Here, we describe 'ExoCET' (Exonuclease Combined with RecET recombination) to directly clone any chosen region from bacterial and mammalian genomes with nucleotide precision into operational plasmids. ExoCET combines in vitro exonuclease and annealing with the remarkable capacity of full length RecET homologous recombination (HR) to retrieve specified regions from genomic DNA preparations...
December 12, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29230873/non-coded-amino-acids-in-protein-engineering-structure-activity-relationship-studies-of-hirudin-thrombin-interaction
#9
Vincenzo De Filippis, Laura Acquasaliente, Giulia Pontarollo, Daniele Peterle
The advent of recombinant DNA technology allowed to site-specifically insert, delete or mutate almost any amino acid in a given protein, significantly improving our knowledge of protein structure, stability and function. Nevertheless, a quantitative description of the physical and chemical basis that makes a polypeptide chain to efficiently fold into a stable and functionally active conformation is still elusive. This mainly originates from the fact that nature combined, in a yet unknown manner, different properties (i...
December 12, 2017: Biotechnology and Applied Biochemistry
https://www.readbyqxmd.com/read/29202040/a-simple-and-universal-system-for-gene-manipulation-in-aspergillus-fumigatus-in-vitro-assembled-cas9-guide-rna-ribonucleoproteins-coupled-with-microhomology-repair-templates
#10
Qusai Al Abdallah, Wenbo Ge, Jarrod R Fortwendel
CRISPR (clustered regularly interspaced short palindromic repeat)-Cas9 is a novel genome-editing system that has been successfully established in Aspergillus fumigatus. However, the current state of the technology relies heavily on DNA-based expression cassettes for delivering Cas9 and the guide RNA (gRNA) to the cell. Therefore, the power of the technology is limited to strains that are engineered to express Cas9 and gRNA. To overcome such limitations, we developed a simple and universal CRISPR-Cas9 system for gene deletion that works across different genetic backgrounds of A...
November 2017: MSphere
https://www.readbyqxmd.com/read/29191048/drug-delivery-vectors-based-on-filamentous-bacteriophages-and-phage-mimetic-nanoparticles
#11
Zhigang Ju, Wei Sun
With the development of nanomedicine, a mass of nanocarriers have been exploited and utilized for targeted drug delivery, including liposomes, polymers, nanoparticles, viruses, and stem cells. Due to huge surface bearing capacity and flexible genetic engineering property, filamentous bacteriophage and phage-mimetic nanoparticles are attracting more and more attentions. As a rod-like bio-nanofiber without tropism to mammalian cells, filamentous phage can be easily loaded with drugs and directly delivered to the lesion location...
November 2017: Drug Delivery
https://www.readbyqxmd.com/read/29184994/synthetic-biology-for-microbial-heavy-metal-biosensors
#12
REVIEW
Hyun Ju Kim, Haeyoung Jeong, Sang Jun Lee
Using recombinant DNA technology, various whole-cell biosensors have been developed for detection of environmental pollutants, including heavy metal ions. Whole-cell biosensors have several advantages: easy and inexpensive cultivation, multiple assays, and no requirement of any special techniques for analysis. In the era of synthetic biology, cutting-edge DNA sequencing and gene synthesis technologies have accelerated the development of cell-based biosensors. Here, we summarize current technological advances in whole-cell heavy metal biosensors, including the synthetic biological components (bioparts), sensing and reporter modules, genetic circuits, and chassis cells...
November 28, 2017: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/29184391/novel-production-method-of-innovative-antiangiogenic-and-antitumor-small-peptides-in-escherichia-coli
#13
Sarra Setrerrahmane, Jian Yu, Jingchao Hao, Heng Zheng, Hanmei Xu
Background: Developing innovative drugs with potent efficacy, specificity, and high safety remains an ongoing task in antitumor therapy development. In the last few years, peptide drugs have become attractive agents in cancer therapy. HM-3, mainly with antiangiogenic effect, and AP25, with an additional antiproliferative effect, are two peptides designed in our laboratory targeting αvβ3 and α5β1 integrins, respectively. The low molecular weight of the two peptides renders their recombinant expression very difficult, and the complicated structure of AP25 makes its chemical synthesis restricted, which presents a big challenge for its development...
2017: Drug Design, Development and Therapy
https://www.readbyqxmd.com/read/29179262/advances-towards-controlling-meiotic-recombination-for-plant-breeding
#14
Kyuha Choi
Meiotic homologous recombination generates new combinations of preexisting genetic variation and is a crucial process in plant breeding. Within the last decade, our understanding of plant meiotic recombination and genome diversity has advanced considerably. Innovation in DNA sequencing technology has led to the exploration of high-resolution genetic and epigenetic information in plant genomes, which has helped to accelerate plant breeding practices via high-throughput genotyping, and linkage and association mapping...
November 23, 2017: Molecules and Cells
https://www.readbyqxmd.com/read/29176835/enhanced-eicosapentaenoic-acid-production-by-a-new-deep-sea-marine-bacterium-shewanella-electrodiphila-mar441t
#15
Jinwei Zhang, J Grant Burgess
Omega-3 fatty acids are products of secondary metabolism, essential for growth and important for human health. Although there are numerous reports of bacterial production of omega-3 fatty acids, less information is available on the biotechnological production of these compounds from bacteria. The production of eicosapentaenoic acid (EPA, 20:5ω3) by a new species of marine bacteria Shewanella electrodiphila MAR441T was investigated under different fermentation conditions. This strain produced a high percentage (up to 26%) of total fatty acids and high yields (mg / g of biomass) of EPA at or below the optimal growth temperature...
2017: PloS One
https://www.readbyqxmd.com/read/29173169/crispr-cas9-gene-editing-from-basic-mechanisms-to-improved-strategies-for-enhanced-genome-engineering-in-vivo
#16
Jayme Salsman, Jean-Yves Masson, Alexandre Orthwein, Graham Dellaire
Targeted genome editing using the CRISPR/Cas9 technology is becoming a major area of research due to its high potential for the treatment of genetic diseases. Our understanding of this approach has expanded in recent years yet several new challenges have presented themselves as we explore the boundaries of this exciting new technology. Chief among these is improving the efficiency but also the preciseness of genome editing. The efficacy of CRISPR/Cas9 technology relies in part on the use of one of the major DNA repair pathways, Homologous recombination (HR), which is primarily active in S and G2 phases of the cell cycle...
November 21, 2017: Current Gene Therapy
https://www.readbyqxmd.com/read/29170960/assembly-and-multiplex-genome-integration-of-metabolic-pathways-in-yeast-using-casemblr
#17
Tadas Jakočiūnas, Emil D Jensen, Michael K Jensen, Jay D Keasling
Genome integration is a vital step for implementing large biochemical pathways to build a stable microbial cell factory. Although traditional strain construction strategies are well established for the model organism Saccharomyces cerevisiae, recent advances in CRISPR/Cas9-mediated genome engineering allow much higher throughput and robustness in terms of strain construction. In this chapter, we describe CasEMBLR, a highly efficient and marker-free genome engineering method for one-step integration of in vivo assembled expression cassettes in multiple genomic sites simultaneously...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29167553/updated-summary-of-genome-editing-technology-in-human-cultured-cells-linked-to-human-genetics-studies
#18
REVIEW
Tatsuo Miyamoto, Silvia Natsuko Akutsu, Shinya Matsuura
Current deep-sequencing technology provides a mass of nucleotide variations associated with human genetic disorders to accelerate the identification of causative mutations. To understand the etiology of genetic disorders, reverse genetics in human cultured cells is a useful approach for modeling a disease in vitro. However, gene targeting in human cultured cells is difficult because of their low activity of homologous recombination. Engineered endonucleases enable enhancement of the local activation of DNA repair pathways at the human genome target site to rewrite the desired sequence, thereby efficiently generating disease-modeling cultured cell clones...
October 11, 2017: Journal of Human Genetics
https://www.readbyqxmd.com/read/29165646/development-and-application-of-a-recombination-based-library-versus-library-high-throughput-yeast-two-hybrid-rll-y2h-screening-system
#19
Fang Yang, Yingying Lei, Meiling Zhou, Qili Yao, Yichao Han, Xiang Wu, Wanshun Zhong, Chenghang Zhu, Weize Xu, Ran Tao, Xi Chen, Da Lin, Khaista Rahman, Rohit Tyagi, Zeshan Habib, Shaobo Xiao, Dang Wang, Yang Yu, Huanchun Chen, Zhenfang Fu, Gang Cao
Protein-protein interaction (PPI) network maintains proper function of all organisms. Simple high-throughput technologies are desperately needed to delineate the landscape of PPI networks. While recent state-of-the-art yeast two-hybrid (Y2H) systems improved screening efficiency, either individual colony isolation, library preparation arrays, gene barcoding or massive sequencing are still required. Here, we developed a recombination-based 'library vs library' Y2H system (RLL-Y2H), by which multi-library screening can be accomplished in a single pool without any individual treatment...
November 20, 2017: Nucleic Acids Research
https://www.readbyqxmd.com/read/29164507/enzyme-linked-immunosorbent-assay-for-the-quantitative-qualitative-analysis-of-plant-secondary-metabolites
#20
REVIEW
Seiichi Sakamoto, Waraporn Putalun, Sornkanok Vimolmangkang, Waranyoo Phoolcharoen, Yukihiro Shoyama, Hiroyuki Tanaka, Satoshi Morimoto
Immunoassays are antibody-based analytical methods for quantitative/qualitative analysis. Since the principle of immunoassays is based on specific antigen-antibody reaction, the assays have been utilized worldwide for diagnosis, pharmacokinetic studies by drug monitoring, and the quality control of commercially available products. Berson and Yalow were the first to develop an immunoassay, known as radioimmunoassay (RIA), for detecting endogenous plasma insulin [1], a development for which Yalow was awarded the Nobel Prize in Physiology or Medicine in 1977...
November 21, 2017: Journal of Natural Medicines
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