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recombinant DNA technology

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https://www.readbyqxmd.com/read/28325290/rna-guided-crispr-cas9-system-mediated-engineering-of-acute-myeloid-leukemia-mutations
#1
Oliver Brabetz, Vijay Alla, Linus Angenendt, Christoph Schliemann, Wolfgang E Berdel, Maria-Francisca Arteaga, Jan-Henrik Mikesch
Current acute myeloid leukemia (AML) disease models face severe limitations because most of them induce un-physiological gene expressions that do not represent conditions in AML patients and/or depend on external promoters for regulation of gene expression/repression. Furthermore, many AML models are based on reciprocal chromosomal translocations that only reflect the minority of AML patients, whereas more than 50% of patients have a normal karyotype. The majority of AML, however, is driven by somatic mutations...
March 17, 2017: Molecular Therapy. Nucleic Acids
https://www.readbyqxmd.com/read/28323062/new-strategies-and-approaches-for-engineering-biosynthetic-gene-clusters-of-microbial-natural-products
#2
REVIEW
Lei Li, Weihong Jiang, Yinhua Lu
With the rapidly growing number of sequenced microbial (meta)genomes, enormous cryptic natural product (NP) biosynthetic gene clusters (BGCs) have been identified, which are regarded as a rich reservoir for novel drug discovery. A series of powerful tools for engineering BGCs has accelerated the discovery and development of pharmaceutically active NPs. Here, we describe recent advances in the strategies for BGCs manipulation, which are driven by emerging technologies, including efficient DNA recombination systems, versatile CRISPR/Cas9 genome editing tools and diverse DNA assembly methods...
March 16, 2017: Biotechnology Advances
https://www.readbyqxmd.com/read/28317025/marker-recycling-in-candida-albicans-through-crispr-cas9-induced-marker-excision
#3
Manning Y Huang, Aaron P Mitchell
We describe here a new approach to marker recycling, a controlled sequence of steps in which a genetic marker is selected and then lost. Marker recycling is important for genetic manipulation, because it allows a single selection marker to be used repeatedly. Our approach relies upon the ability of the CRISPR-Cas9 system to make a targeted double-strand break in DNA and the expectation that a double-strand break within a selection marker may promote recombination between directly repeated sequences that flank the marker...
March 2017: MSphere
https://www.readbyqxmd.com/read/28316014/rescue-of-recombinant-newcastle-disease-virus-a-short-history-of-how-it-all-started
#4
REVIEW
Aidin Molouki, Ben Peeters
Reverse genetics of viruses has come a long way, and many recombinant viruses have been generated since the first successful "rescues" were reported in the late 1970s. Recombinant Newcastle disease virus (rNDV), a non-segmented negative-sense RNA virus (NSNSV), was first rescued in 1999 using a reverse genetics approach similar to that reported for other recombinant viruses of the order Mononegavirales a few years before. The route from an original NDV isolate to the generation of its recombinant counterpart requires many steps that have to be sequentially and carefully completed...
March 18, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28303365/immunoglobulin-like-domain-4-mediated-ligand-independent-dimerization-triggers-vegfr-2-activation-in-huvecs-and-vegfr2-positive-breast-cancer-cells
#5
Sheng Zhang, Xiaoping Gao, Wei Fu, Shengwei Li, Limin Yue
PURPOSE: The extracellular region (EC) of the vascular endothelial growth factor (VEGF) receptor-2 (VEGFR-2) contains seven immunoglobulin-like (Ig-like) domains that are required for specific ligand binding and receptor dimerization. Studies of domain 4-7 deletions and substitutions provided insights into the interaction between receptors in the absence of VEGF. In this study, we investigated the effect of domain 4 in ligand-independent VEGFR-2 dimerization and activation in human vascular endothelial cells and human breast cancer cells...
March 16, 2017: Breast Cancer Research and Treatment
https://www.readbyqxmd.com/read/28283933/designing-of-a-recombinant-agip-bacmid-construct-with-infectious-properties-against-black-cutworm-agrotis-ipsilon-larvae
#6
Naglaa A Abdallah, Wael El-Menofy, Abdelhadi Abdallah Abdelhadi
In this study, Agrotis ipsilon nucleopolyhedrovirus bacmid has been constructed as an infectious bacmid in an attempt to allow genome recombination and generation of virus mutants. Since the FseI, a unique restriction site, is located in a viral coding region (ORF_119), PCR was performed to partially amplify the ORF_119 fragment containing the FseI site to facilitate the bacmid construction in a proper way without interrupting the ORF expression. Construction with repeated fragments at the end of the cloned viral was carried out in an attempt to facilitate circulation during infection in insect cells...
March 10, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28282262/new-insights-for-therapeutic-recombinant-human-mirnas-heterologous-production-rhodovolum-sulfidophilum-vs-escherichia-coli
#7
Patrícia Pereira, Augusto Q Pedro, João A Queiroz, Ana R Figueiras, Fani Sousa
RNA interference-based technologies have emerged as an attractive and effective therapeutic option with potential application in diverse human diseases. These tools rely on the development of efficient strategies to obtain homogeneous non-coding RNA samples with adequate integrity and purity, thus avoiding non-targeted gene-silencing and related side-effects that impair their application onto pre-clinical practice. These RNAs have been preferentially obtained by in vitro transcription using DNA templates or via chemical synthesis...
March 10, 2017: Bioengineered
https://www.readbyqxmd.com/read/28261237/progress-in-genome-editing-technology-and-its-application-in-plants
#8
REVIEW
Kai Zhang, Nadia Raboanatahiry, Bin Zhu, Maoteng Li
Genome editing technology (GET) is a versatile approach that has progressed rapidly as a mechanism to alter the genotype and phenotype of organisms. However, conventional genome modification using GET cannot satisfy current demand for high-efficiency and site-directed mutagenesis, retrofitting of artificial nucleases has developed into a new avenue within this field. Based on mechanisms to recognize target genes, newly-developed GETs can generally be subdivided into three cleavage systems, protein-dependent DNA cleavage systems (i...
2017: Frontiers in Plant Science
https://www.readbyqxmd.com/read/28247205/the-mechanism-of-jurkat-cells-apoptosis-induced-by-aggregatibacter-actinomycetemcomitans-cytolethal-distending-toxin
#9
Hui-Ping Chen, Lu Li, Xu Chen, Mi-Fang Yang, Yu Ye, Xiao-Qian Wang, Yan Xu
Cytolethal distending toxin (CDT) which is produced by Aggregatibacter actinomycetemcomitans causes apoptosis in lymphocytes. But the specific mechanism is not clear. The aim of our research was to investigate the effect and mechanism during this process. The wild-type CdtA, CdtB, CdtC (CdtA(W), CdtB(W), CdtC(W)) and mutant CdtB (CdtB(M)) were expressed and purified respectively and the purity of each subunit was examined by BandScan software. And the type I deoxyribonuclease and PI-3,4,5-triphosphate (PI-3,4,5-P3, PIP3) phosphatase activity were detected by DNA agarose gel electrophoresis and enzyme-linked immunosorbent assay respectively...
February 28, 2017: Apoptosis: An International Journal on Programmed Cell Death
https://www.readbyqxmd.com/read/28242597/advances-in-site-specific-integration-of-transgene-in-animal-genome
#10
Li Guoling, Zhong Cuili, Mo Jianxin, Quan Rong, Wu Zhenfang, Li Zicong, Yang Huaqiang, Zhang Xianwei
The traditional transgenic technologies, such as embryo microinjection, transposon-mediated integration, or lentiviral transfection, usually result in random insertions of the foreign DNA into the host genome, which could have various disadvantages in the establishment of transgenic animals. Therefore, a strategy for site-specific integration of a transgene is needed to generate genetically modified animals with accurate and identical genotypes. However, the efficiency for site-specific integration of transgene is very low, which is mainly caused by two issues...
February 20, 2017: Yi Chuan, Hereditas
https://www.readbyqxmd.com/read/28240182/integration-of-novel-materials-and-advanced-omics-technologies-into-new-vaccine-design
#11
Wenzhen Liao, Tian-Tian Zhang, Liqian Gao, Su Seong Lee, Jie Xu, Han Zhang, Zhaogang Yang, Zhaoyu Liu, Wen Li
Designing new vaccines is one of the most challenging tasks for public health to prevent both infectious and chronic diseases. Even though many research scientists have spent great efforts in improving the specificity, sensitivity and safety of current available vaccines, there are still much space on how to effectively combine different biomaterials and technologies to design universal or personalized vaccines. Traditionally, vaccines were made based on empirical approaches designed to mimic immunity induced by natural infection...
February 24, 2017: Current Topics in Medicinal Chemistry
https://www.readbyqxmd.com/read/28236853/in-pursuit-of-cancer-metastasis-therapy-by-bacteria-and-its-biofilms-history-or-future
#12
Mohd Adnan, Saif Khan, Eyad Al-Shammari, Mitesh Patel, Mohd Saeed, Sibte Hadi
The 20th century observation of increasing comprehensive load of cancer, advanced cancer prevention strategies, creative hypotheses and control procedures by research communities are being traversed and stimulated in multiple facets. Inference of genetically modified non-pathogenic and natural bacterial species as potential anti-tumor agents is one such original perspective. Live, genetically modified non-pathogenic or attenuated bacterial species are able to form biofilms by multiplying selectively or non-selectively on cancer cells, which will lead to metastasis disruption...
March 2017: Medical Hypotheses
https://www.readbyqxmd.com/read/28223401/transcriptome-analyses-of-rhesus-monkey-pre-implantation-embryos-reveal-a-reduced-capacity-for-dna-double-strand-break-repair-in-primate-oocytes-and-early-embryos
#13
Xinyi Wang, Denghui Liu, Dajian He, Shengbo Suo, Xian Xia, Xiechao He, Jing-Dong Han, Ping Zheng
Pre-implantation embryogenesis encompasses several critical events including genome reprogramming, zygotic genome activation (ZGA), and cell fate commitment, most of which remain mechanistically unclear in primates. In addition, primates display a high rate of embryo wastage without any clear molecular basis. Understanding the factors involved in genome reprogramming and ZGA will help the generation of induced pluripotent stem cells with high efficiency. Moreover, explaining the molecular basis responsible for embryo wastage in primates will greatly expand our knowledge of species evolution...
February 21, 2017: Genome Research
https://www.readbyqxmd.com/read/28220435/rapid-construction-of-multiplexed-crispr-cas9-systems-for-plant-genome-editing
#14
Levi Lowder, Aimee Malzahn, Yiping Qi
Multiplex CRISPR-Cas9 nuclease mediated genome editing is an efficient method for disrupting gene function in plants. Use of CRISPR-Cas9 has escalated rapidly in recent years and is expected to become routine practice in molecular biology and related fields of research. Due to the relatively novel and widespread adoption of this technology, first-time users may not have regular access to experienced guidance or technical support from peers or mentors. Here, we offer guidance and technical support in the form of a detailed and tested protocol for simultaneous targeting of three separate loci on the TRANSPARENT TESTA 4 (TT4) gene in Arabidopsis thaliana using multiplex CRISPR-Cas9...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28213165/hbvcircle-a-novel-tool-to-investigate-hepatitis-b-virus-covalently-closed-circular-dna
#15
Zhipeng Yan, Jing Zeng, Youjun Yu, Kunlun Xiang, Hui Hu, Xue Zhou, Lili Gu, Li Wang, Jie Zhao, John A T Young, Lu Gao
BACKGROUND & AIMS: Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) persists as a stable episome in infected hepatocytes and serves as a template for the transcription of all viral genes. Due to the narrow host range of HBV, the development of a robust mouse model that supports cccDNA-dependent viral replication is a key hurdle in the development of novel HBV therapeutics. This study aimed to develop a novel tool to investigate HBV cccDNA. METHODS: Through minicircle technology, HBVcircle, a recombinant cccDNA, was easily generated and extracted from a genetically engineered E...
February 14, 2017: Journal of Hepatology
https://www.readbyqxmd.com/read/28212457/papillomaviruses-a-systematic-review
#16
Rodrigo Pinheiro Araldi, Suely Muro Reis Assaf, Rodrigo Franco de Carvalho, Márcio Augusto Caldas Rocha de Carvalho, Jacqueline Mazzuchelli de Souza, Roberta Fiusa Magnelli, Diego Grando Módolo, Franco Peppino Roperto, Rita de Cassia Stocco, Willy Beçak
In the last decades, a group of viruses has received great attention due to its relationship with cancer development and its wide distribution throughout the vertebrates: the papillomaviruses. In this article, we aim to review some of the most relevant reports concerning the use of bovines as an experimental model for studies related to papillomaviruses. Moreover, the obtained data contributes to the development of strategies against the clinical consequences of bovine papillomaviruses (BPV) that have led to drastic hazards to the herds...
February 16, 2017: Genetics and Molecular Biology
https://www.readbyqxmd.com/read/28209186/identification-of-immune-protective-genes-of-eimeria-maxima-through-cdna-expression-library-screening
#17
XinChao Yang, MengHui Li, JianHua Liu, YiHong Ji, XiangRui Li, LiXin Xu, RuoFeng Yan, XiaoKai Song
BACKGROUND: Eimeria maxima is one of the most prevalent Eimeria species causing avian coccidiosis, and results in huge economic loss to the global poultry industry. Current control strategies, such as anti-coccidial medication and live vaccines have been limited because of their drawbacks. The third generation anticoccidial vaccines including the recombinant vaccines as well as DNA vaccines have been suggested as a promising alternative strategy. To date, only a few protective antigens of E...
February 16, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28191922/synergistic-integration-of-experimental-and-simulation-approaches-for-the-de-novo-design-of-silk-based-materials
#18
Wenwen Huang, Davoud Ebrahimi, Nina Dinjaski, Anna Tarakanova, Markus J Buehler, Joyce Y Wong, David L Kaplan
Tailored biomaterials with tunable functional properties are crucial for a variety of task-specific applications ranging from healthcare to sustainable, novel bio-nanodevices. To generate polymeric materials with predictive functional outcomes, exploiting designs from nature while morphing them toward non-natural systems offers an important strategy. Silks are Nature's building blocks and are produced by arthropods for a variety of uses that are essential for their survival. Due to the genetic control of encoded protein sequence, mechanical properties, biocompatibility, and biodegradability, silk proteins have been selected as prototype models to emulate for the tunable designs of biomaterial systems...
February 13, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28186591/-application-of-recombinant-gp%C3%A2-a-combined-luminex-beads-for-the-detection-of-hpa-1a-antibody
#19
Sudan Tao, Ying Liu, Yanming He, Yanling Ying, Ji He, Faming Zhu
OBJECTIVE: To generate recombinant GPⅢa as an alternative source for HPA-1a antigen and combine it with Luminex xMAP beads for the detection of HPA-1a-specific alloantibody. METHODS: The full coding region of ITGB3 gene was amplified and ligated with pcDNA3.1. The recombinant plasmid was transfected into CHO cells, and those with stable expression were screened with G418. Expressed protein was identified and coupled with Luminex xMAP beads, which were then reacted with sera samples...
February 10, 2017: Zhonghua Yi Xue Yi Chuan Xue za Zhi, Zhonghua Yixue Yichuanxue Zazhi, Chinese Journal of Medical Genetics
https://www.readbyqxmd.com/read/28167937/antiviral-defenses-in-plants-through-genome-editing
#20
REVIEW
Gustavo Romay, Claude Bragard
Plant-virus interactions based-studies have contributed to increase our understanding on plant resistance mechanisms, providing new tools for crop improvement. In the last two decades, RNA interference, a post-transcriptional gene silencing approach, has been used to induce antiviral defenses in plants with the help of genetic engineering technologies. More recently, the new genome editing systems (GES) are revolutionizing the scope of tools available to confer virus resistance in plants. The most explored GES are zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeats/Cas9 endonuclease...
2017: Frontiers in Microbiology
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