1.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
2.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*
3.Functional characterization of piggyBac-like elements from Nilaparvata lugens (Stål) (Hemiptera: Delphacidae).
Jun LYU ; Qin SU ; Jinhui LIU ; Lin CHEN ; Jiawei SUN ; Wenqing ZHANG
Journal of Zhejiang University. Science. B 2022;23(6):515-527
PiggyBac is a transposable DNA element originally discovered in the cabbage looper moth (Trichoplusia ni). The T. ni piggyBac transposon can introduce exogenous fragments into a genome, constructing a transgenic organism. Nevertheless, the comprehensive analysis of endogenous piggyBac-like elements (PLEs) is important before using piggyBac, because they may influence the genetic stability of transgenic lines. Herein, we conducted a genome-wide analysis of PLEs in the brown planthopper (BPH) Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), and identified a total of 28 PLE sequences. All N. lugens piggyBac-like elements (NlPLEs) were present as multiple copies in the genome of BPH. Among the identified NlPLEs, NlPLE25 had the highest copy number and it was distributed on five chromosomes. The full length of NlPLE25 consisted of terminal inverted repeats and sub-terminal inverted repeats at both terminals, as well as a single open reading frame transposase encoding 546 amino acids. Furthermore, NlPLE25 transposase caused precise excision and transposition in cultured insect cells and also restored the original TTAA target sequence after excision. A cross-recognition between the NlPLE25 transposon and the piggyBac transposon was also revealed in this study. These findings provide useful information for the construction of transgenic insect lines.
Amino Acid Sequence
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Animals
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Animals, Genetically Modified
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DNA Transposable Elements/genetics*
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Hemiptera/genetics*
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Transposases/genetics*
4.Bt transgenic crops for insect-resistance and modification of Bt protein and utilization of stacking strategy.
Chinese Journal of Biotechnology 2015;31(1):53-64
Insecticidal protein genes from Bacillus thuringiensis are currently the most widely used insect-resistant genes. They have been transferred to many crops for breeding and production. Among them, cotton, maize, potato and other insect-resistant crops are commercialized, creating considerable economic benefit. In this review, we summarized advances in identifying functional genes and transgenic crops for insect resistance, compared different strategies for enhancing vigor of insecticidal protein and utilizing gene stacking as well as listing valuable groups of stacked genes. In addition, the methods for multiple gene transformation was discussed.
Animals
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Bacterial Proteins
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genetics
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Crops, Agricultural
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genetics
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Endotoxins
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genetics
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Hemolysin Proteins
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genetics
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Insecta
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Plants, Genetically Modified
5.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
6.Preparation of transgenic Musca domestica by microinjection method.
Lanchen WANG ; Yang YANG ; Xiaoli SHANG ; Bing WANG ; Lin YUAN ; Guiming ZHU
Chinese Journal of Biotechnology 2021;37(2):655-662
The transposon vector containing enhanced green fluorescent protein (EGFP) was injected into early housefly (Musca domestica L.) eggs by microinjection method to realize stable gene expression in vivo for verification, and to study housefly gene function. A borosilicate glass micro injection needle suitable for microinjection of housefly eggs was made, the softening treatment conditions of housefly egg shells were explored, and a microinjection technology platform suitable for housefly was constructed with a high-precision microsyringe Nanoject Ⅲ as the main body. The recombinant plasmid PiggyBac-[3×P3]-EGFP containing the eye-specific 3×P3 promoter and EGFP and the stable genetic expression helper plasmid pHA3pig helper were microinjected into the treated housefly eggs. After emergence, the eye luminescence was observed, and the expression and transcription level of EGFP were detected. The results showed that the normal hatching rate of housefly eggs was 55% when rinsed in bleaching water for 35 s. The hardness of the egg shell treated for 35 s was suitable for injection and the injection needle was not easy to break. About 3% of the emerged housefly eyes had green fluorescence. Through further molecular detection, EGFP specific fragments with a size of 750 bp were amplified from DNA and RNA of housefly. Through the technical platform, the stable expression of reporter genes in housefly can be conveniently and effectively realized, and a bioreactor with housefly as the main body can be established, which provides certain reference value for subsequent research on housefly gene function.
Animals
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Animals, Genetically Modified
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Gene Expression
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Genes, Reporter
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Green Fluorescent Proteins/genetics*
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Houseflies/genetics*
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Microinjections
8.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
9.Genetically engineered corn rootworm resistance: potential for reduction of human health effects from pesticides.
Frederick W OEHME ; John A PICKRELL
Biomedical and Environmental Sciences 2003;16(1):17-28
OBJECTIVE AND METHODSInsecticide use, grower preferences regarding genetically engineered (GE) corn resistant to corn rootworm (CRW), and the health effects of using various CRW insecticides (organophosphates, pyrethroids, fipronil and carbamates) are reviewed for current and future farm practices.
RESULTSPest damage to corn has been reduced only one-third by insecticide applications. Health costs from insecticide use appear significant, but costs attributable to CRW control are not quantifiable from available data. Methods reducing health-related costs of insecticide-based CRW control should be evaluated. As a first step, organophosphate insecticide use has been reduced as they have high acute toxicity and risk of long-term neurological consequences. A second step is to use agents which more specifically target the CRW.
CONCLUSIONWhereas current insecticides may be poisonous to many species of insects, birds, mammals and humans, a protein derived from Bacillus thurigiensis and produced in plants via genetic modification can target the specific insect of CRW (Coleoptra), sparing other insect and non-insect species from injury.
Animals ; Consumer Product Safety ; Food, Genetically Modified ; Insecta ; Insecticide Resistance ; Insecticides ; adverse effects ; Pest Control, Biological ; economics ; methods ; Plant Roots ; Plants, Genetically Modified ; Zea mays ; genetics ; growth & development
10.Efficient packaging retrovirus and construction of transgenic chicken technical platform.
Chaolai MAN ; Qing ZHANG ; Yan CHEN ; Dahai ZHU
Journal of Biomedical Engineering 2007;24(5):1111-1117
Transgenic chicken and oviduct bioreactor are growing to be one of the hotspot of scientific study in the field of biology. The most successful method of producing transgenic chicken is pseudotyped retrovirus vector system, but no one has reported the production of transgenic chicken by retrovirus system recently in our country. In order to accelerate our study in this field, we introduced the relevant technical methods such as packaging retrovirus and vesicular stomatitis virus G glycoprotein (VSV-G) pseudotyped retrovirus, optimizing the conditions of packaging retrovirus, concentrating VSV-G pseudotyped retrovirus, helper virus assays, and microinjection of retrovirus. Furthermore, we successfully conducted in vivo study for detecting the marker gene EGFP of chicken embryo as well as in vitro study for detecting that gene of chicken embryo myoblast (CFM), thus we have provided an applied technical platform for studies of transgenic chicken in the future.
Animals
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Animals, Genetically Modified
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Chick Embryo
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Chickens
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genetics
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DNA Primers
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Genetic Vectors
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genetics
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Retroviridae
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genetics
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Vesicular stomatitis Indiana virus
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genetics