1.Retrospect and prospect of transgenic fish breeding in China.
Chinese Journal of Biotechnology 2016;32(7):851-860
The first transgenic fish was generated in China about 30 years ago. Since then, considerable progress has been achieved for farmed fishes breeding with improvement of target traits of growth, disease resistance, stress tolerance, and nutrition qualities. Up to now, the technology of transgenic fish breeding is almost mature and the biosafety assessment is established. In this review, a successful example of the fast-growing transgenic common carp was presented and the foreground of transgenic fish breeding was also discussed and prospected.
Animals
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Animals, Genetically Modified
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Breeding
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China
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Fishes
2.The in vivo imaging of zebrafish for chemical vascular toxicity assessment.
Di JI ; Qi Cheng ZHU ; Yan Hong WEI
Chinese Journal of Preventive Medicine 2022;56(1):33-37
Objective: To evaluate the vascular toxicity of chemicals by a real-time observation approach using the transgenic zebrafish. Methods: The spatiotemporal vascular alterations of transgenic zebrafish after chemical exposure were assessed by laser confocal microscopy and high-content screening analysis, respectively. Results: The method using Laser Confocal Microscopy (LCM) is easier to operate and yields high-resolution images, while it is lower throughput and inefficient. In contrast, high-content analysis (HCA) analysis obtains high-quality data of vascular toxicity manifesting whole blood vasculature, whereas it requires delicate operation procedures and advanced experimental conditions. Conclusion: Two kinds of zebrafish imaging methods each have advantages and disadvantages. LCM is suitable for the evaluation of a small number of chemicals. HCA, a cutting-edge technology, has great potential for chemical safety assessment allowing high throughput vascular toxicity tests of a good number of chemicals at a time.
Animals
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Animals, Genetically Modified
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Cardiovascular System
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Toxicity Tests
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Zebrafish
3.Advances in tumor-therapy using genetically modified Salmonella.
Kang LIANG ; Xinxin ZHAO ; Jie YI ; Qiong LIU ; Qing LIU ; Qingke KONG
Chinese Journal of Biotechnology 2016;32(5):565-576
Tumor is a neoplasm formed by the abnormal proliferation of local tissue cells under the effects of different tumorigenic factors. Tumor-therapy has always been a difficult clinical issue, while regular cancer treatments, such as radiotherapy, chemotherapy and surgery, have obvious limitations. Earlier studies have shown that some obligate anaerobes or facultative anaerobes have anti-tumor effects, for example, Salmonella typhymurium as facultative anaerobic bacteria can selectively colonize tumors and inhibit their growth. Besides, Salmonella has many advantages in tumor-therapy. In the past decade or two, many researchers have carried out genetic manipulation to attenuate the virulence of Salmonella, to improve their specificity of tumor colonization and specially to use attenuated Salmonella as carriers to deliver a variety of anti-tumor therapeutic molecules, and these genetically modified Salmonella have shown good anti-tumor effects in many animal experiments. Along with further research of Salmonella-mediated antitumor treatment, applications of genetically modified Salmonella for more effective tumor-therapy are promising. We reviewed the anti-tumor mechanisms of Salmonella, the research progress in tumor-therapy using genetically modified Salmonella, and current problems and possible solutions.
Animals
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Humans
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Microorganisms, Genetically-Modified
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Neoplasms
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therapy
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Salmonella
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genetics
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Virulence
4.Construction of "lock-key" biological living material based on double engineered bacteria and its application on intestinal retention in vivo.
Minghui ZHANG ; Yingying ZHANG ; Pengcheng ZHAO ; Hanjie WANG
Chinese Journal of Biotechnology 2023;39(3):1163-1174
At present, the research of biological living materials mainly focuses on applications in vitro, such as using a single bacterial strain to produce biofilm and water plastics. However, due to the small volume of a single strain, it is easy to escape when used in vivo, resulting in poor retention. In order to solve this problem, this study used the surface display system (Neae) of Escherichia coli to display SpyTag and SpyCatcher on the surface of two strains, respectively, and constructed a double bacteria "lock-key" type biological living material production system. Through this force, the two strains are cross-linked in situ to form a grid-like aggregate, which can stay in the intestinal tract for a longer time. The in vitro experiment results showed that the two strains would deposit after mixing for several minutes. In addition, confocal imaging and microfluidic platform results further proved the adhesion effect of the dual bacteria system in the flow state. Finally, in order to verify the feasibility of the dual bacteria system in vivo, mice were orally administrated by bacteria A (p15A-Neae-SpyTag/sfGFP) and bacteria B (p15A-Neae-SpyCatcher/mCherry) for three consecutive days, and then intestinal tissues were collected for frozen section staining. The in vivo results showed that the two bacteria system could be more detained in the intestinal tract of mice compared with the non-combined strains, which laid a foundation for further application of biological living materials in vivo.
Animals
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Mice
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Bacteria
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Microorganisms, Genetically-Modified
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Escherichia coli/genetics*
5.Agricultural biotechnology safety assessment.
Scott MCCLAIN ; Wendelyn JONES ; Xiaoyun HE ; Gregory LADICS ; Andrew BARTHOLOMAEUS ; Alan RAYBOULD ; Petra LUTTER ; Haibin XU ; Xue WANG
Chinese Journal of Preventive Medicine 2015;49(1):78-87
Genetically modified (GM) crops were first introduced to farmers in 1995 with the intent to provide better crop yield and meet the increasing demand for food and feed. GM crops have evolved to include a thorough safety evaluation for their use in human food and animal feed. Safety considerations begin at the level of DNA whereby the inserted GM DNA is evaluated for its content, position and stability once placed into the crop genome. The safety of the proteins coded by the inserted DNA and potential effects on the crop are considered, and the purpose is to ensure that the transgenic novel proteins are safe from a toxicity, allergy, and environmental perspective. In addition, the grain that provides the processed food or animal feed is also tested to evaluate its nutritional content and identify unintended effects to the plant composition when warranted. To provide a platform for the safety assessment, the GM crop is compared to non-GM comparators in what is typically referred to as composition equivalence testing. New technologies, such as mass spectrometry and well-designed antibody-based methods, allow better analytical measurements of crop composition, including endogenous allergens. Many of the analytical methods and their intended uses are based on regulatory guidance documents, some of which are outlined in globally recognized documents such as Codex Alimentarius. In certain cases, animal models are recommended by some regulatory agencies in specific countries, but there is typically no hypothesis or justification of their use in testing the safety of GM crops. The quality and standardization of testing methods can be supported, in some cases, by employing good laboratory practices (GLP) and is recognized in China as important to ensure quality data. Although the number of recommended, in some cases, required methods for safety testing are increasing in some regulatory agencies, it should be noted that GM crops registered to date have been shown to be comparable to their nontransgenic counterparts and safe . The crops upon which GM development are based are generally considered safe.
Agriculture
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Animal Feed
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Animals
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Biotechnology
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China
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Consumer Product Safety
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Food, Genetically Modified
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Humans
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Models, Animal
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Plants, Genetically Modified
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Safety
6.Biosafety assessment of genetically engineered animals: a review.
Chinese Journal of Biotechnology 2012;28(3):267-281
With the research and development of genetically engineered animals (GEAs) in breeding of new variety, xenotransplantation, bioreactor and disease model, biosafety issues of GEAs have attracted widespread attentions worldwide. So far, governments and agencies have established corresponding laws and regulations to regulate research and application of GEAs or their derived products. We reviewed research contents, evaluated principles, policies and procedures for biosafety of GEAs, also enumerated upcoming approved products of GEAs. Finally, we suggested perspectives of research and application of GEAs or their derived products.
Agriculture
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economics
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legislation & jurisprudence
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Animals
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Animals, Genetically Modified
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genetics
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growth & development
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Biotechnology
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economics
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legislation & jurisprudence
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Food Safety
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Food, Genetically Modified
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economics
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standards
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Laboratories
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standards
7.Advances in spermatogonial transplantation.
National Journal of Andrology 2003;9(1):58-66
Spermatogonial transplantation (ST) is a novel technique which mechanism is similar to other transplantations. The donor testicular cells transferred into recipient testes by the microinjection can initiate spermatogenesis and produce sperm in the recipient testis. The homograft has got success. The xenograft in which (man, horse, bull or rat, can be the donor; immunodeficiency mouse can be the recipient) and cryopreserved germ cells can not clonize in the recipient testes. The reason may be immunoreaction. ST technique will be propitious to these fields, 1. to investigate fundamental aspects of spermatogenesis; 2. to regenerate spermatogenesis in infertile individuals; 3. to develop transgenic animals.
Animals
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Animals, Genetically Modified
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Humans
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Male
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Spermatogenesis
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physiology
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Spermatogonia
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transplantation
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Transplantation, Heterologous
8.Retinol dehydrogenase, RDH1l, is essential for the heart development and cardiac performance in zebrafish.
Wei WANG ; Li-feng ZHANG ; Yong-hao GUI ; Hou-yan SONG
Chinese Medical Journal 2013;126(4):722-728
BACKGROUNDRetinoic acid (RA) is a potent signaling molecule that plays pleiotropic roles in patterning, morphogenesis, and organogenesis during embryonic development. The synthesis from retinol (vitamin A) to retinoic acid requires two sequential oxidative steps. The first step involves the oxidation of retinol to retinal through the action of retinol dehydrogenases. Retinol dehydrogenases1l (RDH1l) is a novel zebrafish retinol dehydrogenase. Herein we investigated the role of zebrafish RDH1l in heart development and cardiac performance in detail.
METHODSRDH1l specific morpholino was used to reduce the function of RDH1l in zebrafish. The gene expressions were observed by using whole mount in situ hybridization. Heart rates were observed and recorded under the microscope from 24 to 72 hours post fertilization (hpf). The cardiac performance was analyzed by measuring ventricular shortening fraction (VSF).
RESULTSThe knock-down of RDH1l led to abnormal neural crest cells migration and reduced numbers of neural crest cells in RDH1l morphant embryos. The reduced numbers of cardiac neural crest cells also can be seen in RDH1l morphant embryos. Furthermore, the morpholino-mediated knock-down of RDH1l resulted in the abnormal heart loop. The left-right determining genes expression pattern was altered in RDH1l morphant embryos. The impaired cardiac performance was observed in RDH1l morphant embryos. Taken together, these data demonstrate that RDH1l is essential for the heart development and cardiac performance in zebrafish.
CONCLUSIONSRDH1l plays a important role in the neural crest cells development, and then ultimately affects the heart loop and cardiac performance. These results show for the first time that an enzyme involved in the retinol to retinaldehyde conversion participate in the heart development and cardiac performance in zebrafish.
Alcohol Oxidoreductases ; genetics ; metabolism ; Animals ; Animals, Genetically Modified ; Heart ; embryology ; Zebrafish ; Zebrafish Proteins ; genetics ; metabolism
9.Transgenic animals bioreactors.
Ke-Mian GOU ; Xiao-Rong AN ; Jian-Hui TIAN ; Yong-Fu CHEN
Chinese Journal of Biotechnology 2002;18(2):144-148
The production of human recombinant proteins in milk of transgenic farm animals offers a safe, very cost-effective source of commercially important proteins that cannot be produced as efficiently in adequate quantities by other methods. This review has summarized the current status of gene selection, vector construct, transgenic methods, economics, and obvious potential in transgenic animals bioreactors. Recently, a more powerful approach was adopted in the transgenic animals founded on the application of nuclear transfer. As we will illustrate, this strategy presents a breakthrough in the overall efficiency of generating transgenic farm animals, product consistency, and time of product development. The successful adaptation of Cre-/lox P-mediated site-specific DNA recombination systems in farm animals will offer unprecedented possibilities for generating transgenic animals.
Animals
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Animals, Genetically Modified
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Bioreactors
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Breast
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metabolism
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Cell Transplantation
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Gene Expression
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Humans
10.Increasing transgenic expression in recombinant Chinese hamster ovary cells using introns in different directions.
Weihua DONG ; Cuiping LI ; Yun YANG ; Tianyun WANG ; Fang WANG
Chinese Journal of Biotechnology 2019;35(6):1071-1078
The aim of this study is to investigate the effect of the chimeric intron in different directions on the expression of the nerve growth factor (NGF) in recombinant Chinese hamster ovary (CHO) cells. The chimeric intron that contained the splice sequence of the first intron of the human β-globin and the human immunoglobulin heavy chain variable region intron was used. NGF gene was cloned into the expression vectors containing the chimeric intron in the forward or reverse direction, followed by transfecting into CHO cells, and screened under G418 to produce the stable transfected CHO cells. Fluorescence quantitative PCR, ELISA, and Western blotting were performed to detect the recombinant NGF gene expression in CHO cells. The results showed that the chimeric introns could significantly enhance the expression of NGF in recombinant CHO cells. Moreover, the enhancing effect on NGF expression level by the intron in the forward direction showed stronger than that of the reverse direction both at mRNA and protein level. In conclusion, the chimeric intron could increase NGF expression in stably transfected CHO cells and the effect is associated with the direction of the intron insertion.
Animals
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Animals, Genetically Modified
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CHO Cells
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Cricetinae
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Cricetulus
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Gene Expression
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Humans
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Introns
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Transfection