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Transgenic Research

Do Van Giap, Jae-Wan Jung, Nan-Sun Kim
Cyclic citrullinated peptide (CCP) antibody has been shown recently to be a promising marker for early detection and diagnosis of rheumatoid arthritis (RA). In order to exploit newly developed therapies for RA, early intervention is crucial in preventing irreversible joint damage. Here, we describe use of a plant expression system to produce a CCP antibody that could be used in the early diagnosis of RA. Heavy and light chain gene sequences of a CCP monoclonal antibody (CCP mAb) were cloned from the hybridoma cell (12G1) and introduced into two separate plant expression vectors under the control of the rice α-amylase 3D (RAmy3D) promoter system...
February 11, 2019: Transgenic Research
Li-Zeng Guan, Shuai Zhao, Gang Shu, Qing-Yan Jiang, Geng-Yuan Cai, Zhen-Fang Wu, Qian-Yun Xi, Yong-Liang Zhang
Producing heterologous enzymes in the animal digestive tract to improve feed utilization rate is a new research strategy by transgenic technology. In this study, transgenic pigs specifically expressing β-glucanase gene in the intestine were successfully produced by somatic cell nuclear transfer technology in order to improve digestibility of dietary β-glucan and absorption of nutrients. The β-glucanase activity in the intestinal juice of 4 transgenic pigs was found to be 8.59 ± 2.49 U/mL. The feeding trial results showed that the crude protein digestion of 4 transgenic pigs was significantly increased compared with that of the non-transgenic pigs...
January 29, 2019: Transgenic Research
Francisco Ayala, Griselda V Fedrigo, Moises Burachik, Patricia V Miranda
Wheat is the most widely grown cereal grain, occupying a significant portion of the total cultivated land. As drought is the major environmental stressor affecting crop production, yield maintenance under water deficit conditions appears as a highly desirable phenotype for crop improvement. The HaHB4 (Helianthus annuus homeobox 4) gene from sunflower encodes for a transcription factor involved in tolerance to environmental stress. The introduction of HaHB4 in wheat led to the development of event IND-ØØ412-7 (HB4® wheat), which displayed higher yield in production environments of low productivity potential...
January 17, 2019: Transgenic Research
Mao Zhang, Gengyuan Cai, Enqing Zheng, Guangguang Zhang, Yang Li, Zicong Li, Huaqiang Yang, Zhenfang Wu
Xylan is one of the main anti-nutritional factors in pig's feed. Although supplementation of β-xylanase in diet can improve the utilization of nutrients in animals, it is limited by feed cost, manufacturing process and storage stability. To determine whether the expression of endogenous β-xylanase gene xynB in vivo can improve digestibility of dietary xylan and absorption of nutrients, we produced transgenic pigs which express the xynB from Aspergillus Niger CGMCC1067 in the parotid gland via nuclear transfer...
January 14, 2019: Transgenic Research
Tahira Hussain, Emre Aksoy, Mehmet Emin Çalışkan, Allah Bakhsh
Most of the commercialized insect resistant transgenic crops express cry gene(s) isolated from Bacillus thuringiensis; however, intensive cultivation of Bt crops over almost two decades has been questioned regarding its sustainability and durability in pest management. The present study focused on silencing of highly specific molting-associated Ecdysone receptor (EcR) gene of Colorado potato beetle (CPB) using RNA interference (RNAi) approach. The partial cDNA of EcR gene of CPB was amplified using specific primers in sense and anti-sense orientations, and cloned in pRNAi-GG vector flanked by an intronic sequence (pdk)...
January 3, 2019: Transgenic Research
Geon A Kim, Eun Mi Lee, Bumrae Cho, Zahid Alam, Su Jin Kim, Sanghoon Lee, Hyun Ju Oh, Jong Ik Hwang, Curie Ahn, Byeong Chun Lee
Herein, we successfully generated transgenic pigs expressing both soluble human tumor necrosis factor receptor I IgG1 -Fc (shTNFRI-Fc) and human hemagglutinin (HA)-tagged-human heme oxygenase-1 (hHO-1) without Gal epitope. Healthy cloned pigs were produced by somatic cell nuclear transfer (SCNT) using the genetically modified cells. The genetic disruption of the GGTA1 genes and absence of expression of BS-IB4 lectin in tail-derived fibroblast of the SCNT-generated piglets were successfully confirmed. The expression of shTNFRI-Fc and HAhHO-1 was fully identified with protective effect against oxidative stress and apoptosis stimulation...
December 14, 2018: Transgenic Research
Xiangdong Yang, Lu Niu, Wei Zhang, Hongli He, Jing Yang, Guojie Xing, Dongquan Guo, Qianqian Zhao, Xiaofang Zhong, Haiyun Li, Qiyun Li, Yingshan Dong
Viruses constitute a major constraint to soybean production worldwide and are responsible for significant yield losses every year. Although varying degrees of resistance to specific viral strains has been identified in some soybean genetic sources, the high rate of mutation in viral genomes and mixed infections of different viruses or strains under field conditions usually hinder the effective control of viral diseases. In the present study, we generated transgenic soybean lines constitutively expressing the double-strand RNA specific ribonuclease gene PAC1 from Schizosaccharomyces pombe to evaluate their resistance responses to multiple soybean-infecting virus strains and isolates...
December 1, 2018: Transgenic Research
Xiaofeng Liu, Hongbo Liu, Min Wang, Ruiqiang Li, Jianhua Zeng, Delin Mo, Peiqing Cong, Xiaohong Liu, Yaosheng Chen, Zuyong He
Insulin-like growth factor 2 (IGF2) plays an important role in the development of the foetus and in post-natal growth and development. A SNP within intron 3 of porcine IGF2 disrupts a binding site for the repressor, zinc finger BED-type containing 6 (ZBED6), leading to up-regulation of IGF2 in skeletal muscle and major effects on muscle growth, heart size, and fat deposition. This favourable mutation is common in Western commercial pig populations, but is not present in most indigenous Chinese pig breeds. Here, we described the efficient disruption of the ZBED6 binding site motif in intron 3 of IGF2 by CRISPR/Cas9 in porcine embryonic fibroblasts (PEFs) from the indigenous Chinese pig breed, Liang Guang Small Spotted pig...
November 28, 2018: Transgenic Research
Magdalena Wróbel-Kwiatkowska, Mateusz Kropiwnicki, Jacek Żebrowski, Athanasios Beopoulos, Lucyna Dymińska, Jerzy Hanuza, Waldemar Rymowicz
The high demand for new biomaterials makes synthesis of polyhydroxyalkanoates (PHA) in plants an interesting and desirable achievement. Production of polymers in plants is an example of application of biotechnology for improving the properties of plants, e.g. industrial properties, but it can also provide knowledge about plant physiology and metabolism. The subject of the present study was an industrially important plant: flax, Linum usitatissimum L., of a fibre cultivar (cv Nike). In the study the gene encoding PHA synthase from Pseudomonas aeruginosa, fused to a peroxisomal targeting signal, was expressed in flax plants with the aim of modifying the mechanical properties of plants...
November 27, 2018: Transgenic Research
Masashi Numata, R I Klein Geltink, Gerard C Grosveld
The ETS transcription factor ETV7 has been characterized as a hematopoietic oncoprotein, which requires cooperating mutations for its leukemogenic activity. Although the ETV7 gene is highly conserved among vertebrates, part of the rodents, including Mus musculus, deleted the Etv7 gene locus. Many human hematopoietic malignancies upregulate ETV7 expression but contrary to ETV7's role in oncogenesis, its physiological role in normal tissues is unknown. To determine the physiological function of ETV7 in vivo and determine its role in tumorigenesis in a mouse model, we have generated an ETV7 transgenic mouse that carries a single copy of human BAC DNA containing the ETV7 gene locus and its regulatory sequences...
November 27, 2018: Transgenic Research
Xiangdong Yang, Jing Yang, Yisheng Wang, Hongli He, Lu Niu, Dongquan Guo, Guojie Xing, Qianqian Zhao, Xiaofang Zhong, Li Sui, Qiyun Li, Yingshan Dong
Sclerotinia stem rot (SSR), caused by the oxalate-secreting necrotrophic fungal pathogen Sclerotinia sclerotiorum, is one of the devastating diseases that causes significant yield loss in soybean (Glycine max). Until now, effective control of the pathogen is greatly limited by a lack of strong resistance in available commercial soybean cultivars. In this study, transgenic soybean plants overexpressing an oxalic acid (OA)-degrading oxalate oxidase gene OXO from wheat were generated and evaluated for their resistance to S...
November 26, 2018: Transgenic Research
Kusumika Mukherjee, Eric C Liao
Zebrafish transgenic lines provide valuable insights into gene functions, cell lineages and cell behaviors during development. Spatiotemporal control over transgene expression is a critical need in many experimental approaches, with applications in loss- and gain-of-function expression, ectopic expression and lineage tracing experiments. The Cre/loxP recombination system is a powerful tool to provide this control and the demand for validated Cre and loxP zebrafish transgenics is high. One of the major challenges to widespread application of Cre/loxP technology in zebrafish is comparatively small numbers of established tissue-specific Cre or CreERT2 lines...
December 2018: Transgenic Research
Nagesh Sardesai, Stephen Foulk, Wei Chen, Huixia Wu, Emily Etchison, Manju Gupta
Agrobacterium-mediated transformation is a complex process that is widely utilized for generating transgenic plants. However, one of the major concerns of this process is the frequent presence of undesirable T-DNA vector backbone sequences in the transgenic plants. To mitigate this deficiency, a ternary strain of A. tumefaciens was modified to increase the precision of T-DNA border nicking such that the backbone transfer is minimized. This particular strain supplemented the native succinamopine VirD1/VirD2 of EHA105 with VirD1/VirD2 derived from an octopine source (pTi15955), the same source as the binary T-DNA borders tested here, residing on a ternary helper plasmid containing an extra copy of the succinamopine VirB/C/G operons and VirD1...
December 2018: Transgenic Research
Cia-Hin Lau, Yousin Suh
The rapid advancement of CRISPR technology has enabled targeted epigenome editing and transcriptional modulation in the native chromatin context. However, only a few studies have reported the successful editing of the epigenome in adult animals in contrast to the rapidly growing number of in vivo genome editing over the past few years. In this review, we discuss the challenges facing in vivo epigenome editing and new strategies to overcome the huddles. The biggest challenge has been the difficulty in packaging dCas9 fusion proteins required for manipulation of epigenome into the adeno-associated virus (AAV) delivery vehicle...
December 2018: Transgenic Research
Marcela Vilarino, Fabian Patrik Suchy, Sheikh Tamir Rashid, Helen Lindsay, Juan Reyes, Bret Roberts McNabb, Talitha van der Meulen, Mark O Huising, Hiromitsu Nakauchi, Pablo Juan Ross
The production of knock-out (KO) livestock models is both expensive and time consuming due to their long gestational interval and low number of offspring. One alternative to increase efficiency is performing a genetic screening to select pre-implantation embryos that have incorporated the desired mutation. Here we report the use of sheep embryo biopsies for detecting CRISPR/Cas9-induced mutations targeting the gene PDX1 prior to embryo transfer. PDX1 is a critical gene for pancreas development and the target gene required for the creation of pancreatogenesis-disabled sheep...
December 2018: Transgenic Research
Nivedita Seshadri, Sumit Sandhu, Xiaoli Wu, Wenjun Liu, Hao Ding
Regulator of telomere length 1 (RTEL1) DNA helicase has been demonstrated to be essential for the maintenance of telomeres and genomic stability. This function of RTEL1 could be required for protecting stem cells from genomic mutations as suggested by its selective expression in stem cell-zones, as well as by RTEL1 mutations identified in Hoyeraal-Hreidarsson syndrome, a severe dyskeratosis congenita that targets primarily stem cell compartments. As a first step to establish a role of RTEL1 in stem cells, we generated an Rtel1CreERT2 mouse allele in which a tamoxifen-inducible Cre (CreERT2) cDNA was specifically knocked into the Rtel1 genomic locus and controlled by the endogenous Rtel1 regulatory elements...
December 2018: Transgenic Research
Parimala Chinnadurai, Duška Stojšin, Kang Liu, Gregory E Frierdich, Kevin C Glenn, Tao Geng, Adam Schapaugh, Keguo Huang, Andrew E Deffenbaugh, Zi L Liu, Luis A Burzio
The expression of the CP4 EPSPS protein in genetically engineered (GE) soybean confers tolerance to the Roundup® family of agricultural herbicides. This study evaluated the variability of CP4 EPSPS expression using an enzyme-linked immunosorbent assay in soybean tissues collected across diverse germplasm and 74 different environments in Argentina, Brazil and the USA. Evaluated material included single and combined (stacked) trait products with other GE traits in entries with cp4 epsps gene at one or two loci...
December 2018: Transgenic Research
Xiaoxi Wei, Min Hu, Fei Liu
Reporter genes play important roles in transgenic research. LacZ is a widely used reporter gene that encodes Escherichia coli β-galactosidase, an enzyme that is well known for its ability to hydrolyze X-gal into a blue product. It is unknown whether transgenic LacZ has any adverse effects. R26R reporter mice, containing a LacZ reporter gene, were generated to monitor the in vivo recombination activity of various transgenic Cre recombinase via X-gal staining. P0-Cre is expressed in neural crest-derived cells, which give rise to the majority of the craniofacial bones...
December 2018: Transgenic Research
Anik Luke Dhanaraj, Alan Ray Willse, S P Kamath
Bollgard-II cotton expressing Cry1Ac and Cry2Ab2 insecticidal proteins has been commercially cultivated in India since 2006 to control bollworms. These genes were introgressed into parental germplasm of numerous hybrids. Therefore, it is imperative that these insecticidal proteins are expressed in sufficient quantities in different tissues, throughout the season irrespective of genetic background or environmental conditions for effective performance. Here, we document results of a comprehensive study on pattern of expression of Bt proteins across different stages of crop growth in > 2000 cotton hybrids (Gossypium hirsutum), across 12 cropping seasons tested in the Northern, Southern or Central zones in India, in terminal leaf, pre-candle square and boll epicarp tissues...
October 30, 2018: Transgenic Research
Caitlin A Cooper, Mark L Tizard, Tamsyn Stanborough, Sean C Moore, P Scott Chandry, Kristie A Jenkins, Terry G Wise, Terri E O'Neil, Daniel S Layton, Kirsten R Morris, Robert J Moore, Narelle Fegan, Timothy J Doran
Zoonotic and foodborne diseases pose a significant burden, decreasing both human and animal health. Modifying chickens to overexpress antimicrobials has the potential to decrease bacterial growth on poultry products and boost chicken innate immunity. Chickens overexpressing either ovotransferrin or avian β-defensin-3 (AvβD3) were generated using Tol-2 transposons. Transgene expression at the RNA and protein level was seen in egg white, breast muscle, and serum. There were significant differences in the immune cell populations in the blood, bursa, and spleen associated with transgene expression including an increased proportion of CD8+ cells in the blood of ovotransferrin and AvβD3 transgenic birds...
October 29, 2018: Transgenic Research
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