• Development and Reproduction
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• v.23 no.4
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• pp.385-390
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• 2019

Upon gene inactivation in animal models, the zebrafish (Danio rerio) has become a useful model organism for many reasons, including the fact that it is amenable to various forms of genetic manipulation. Genome editing is a type of genetic engineering in which DNA is inserted, deleted, modified, or replaced in the genome of a living organism. Mainly, CRISPR (clustered regularly interspaced short palindromic repeats) Cas9 (CRISPR-associated protein 9) is a technology that enables geneticists to edit parts of the genome. In this study, we utilized this technology to generate an mmp15b mutant by using zebrafish as an animal model. MMP15 is the membrane-type MMP (MT-MMP) which is a recently identified matrix metalloproteinase (MMP) capable of degrading all kinds of extracellular matrix proteins as well as numerous bioactive molecules. Although the newly-established mmp15b zebrafish mutant didn't exhibit morphological phenotypes in the developing embryos, it might be further utilized to understand the role of MMP15 in liver-related diseases, such as liver fibrosis, and associated pathogeneses in humans.

• Korean Journal of Agricultural Science
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• v.45 no.1
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• pp.28-37
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• 2018

This study was done to develop environmental risk assessments and a biosafety guide for insect-resistant genetically modified rice at a LMO (Living Modified Organism) isolation field. In the LMO quarantine area of Kyungpook National University, the species diversities and population densities of non-target insects found on insect-resistant genetically modified rice (Bt-9) resistant to Cnaphalocrocis medinalis and on non-GM rices (Dongjin and Ilmi) were investigated. The Bt-9 event was therefore evaluated under field conditions to detect possible impacts on the above ground insects and spiders. The study compared transgenic rice and two non-GM reference rices, Ilmi and Dongjin, at Gunwi in Southern Korea in 2016. Each rice was grown on three $18m^2$ plots with a randomized block design. A total of 4,243 individuals from 43 families and 9 orders were collected from the LMO isolation field. In the three types of rice fields, a total of 1,467 individuals from the insect-resistant genetically modified rice (Bt-9), 1,423 individuals from the Ilmi, and 1,353 individuals from the Dongjin were collected, respectively. There was no difference between the population densities of the non-target insect pests, natural enemies and other insects on the insect-resistant genetically modified rice (Bt-9) and non-GM rices. These results provide the diversity and population density of non-target insects for an environment risk assessment survey on insect-resistant genetically modified rice and could be used as a guideline to make a biosafety assessment method for genetically modified crops.

• Journal of Applied Biological Chemistry
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• v.61 no.4
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• pp.305-314
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• 2018

'New Breeding Techniques (NBTs)' have been one of hot issues, since their future will be affected profoundly by national as well as international regulatory landscapes. In this review, we compare characteristics of NBTs with conventional and genetic modification, and analyze genetically modified organism (GMO) regulatory systems in the context of possible regulation of NBTs. NBTs are very heterogeneous in terms of principles, methodologies, and final products. As Living Modified Organisms (LMO) is defined in the Cartagena Protocol on Biosafety (CPB) as an organism containing novel combination of genetic materials obtained by the use of modern biotechnology, CPB as well as other national legislations locate itself somewhere in the middle between product-based and process-based regulations. It is also noted that jurisdictions with regulatory systems more oriented to product-based one tend to be more productive and decide or may decide to exempt site-directed nucleases-1 from GMO regulation. In this context, Korean legislations are reviewed to clarify the commons and differences in GMO definitions. Act on Transboundary Movement of LMO Act, Food Sanitation Act and Agricultural and Fishery Products Quality Control Act are three major acts to regulate GMOs. It is noted that there are differences in the definition of LMO or GM food/products especially between the LMO Act and the Food Sanitation Act. Such differences may cause conflicts between Acts when policy-decision regarding the regulation of NBTs is made. Therefore, it is necessary to reorganize legislations before policies regarding the regulation of any techniques from biotechnology are made.

• Korean Journal of Agricultural Science
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• v.48 no.4
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• pp.875-890
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• 2021

In order to confirm the safety of a genetically modified organism (GMO), we assess its potential toxicity on non-target insects and spiders. In this study, the effects of GM soybean, a type of vitamin-A-enhanced transgenic soybean with tolerance to the herbicide glufosinate, were assessed under a field condition. The study compared this vitamin-A-enhanced transgenic soybean and a non-GM soybean (Gwangan) in a living modified organism (LMO) isolated field of Kyungpook National University (Gunwi) and the National Institute Agricultural Sciences (Jeonju) in the Republic of Korea in 2019 - 2020. In total, 207,760 individual insects and arachnids, representing 81 families and 13 orders, were collected during the study. From the two types of soybean fields, corresponding totals of 105,765 and 101,995 individuals from the vitamin-A-enhanced transgenic soybean and Gwangan samples areas were collected. An analysis of variance indicated no significant differences (p < 0.05). A multivariate analysis showed that the dominance and richness outcomes of plant-dwelling insects were similar. The data on insect species population densities were subjected to a principal component analysis (PCA) and an orthogonal partial least squares-discriminant analysis (OPLS-DA), which did not distinguish between the two varieties, i.e., the vitamin-A-enhanced transgenic soybean and the non-GM soybean in any cultivated field. However, the results of the PCA analysis could be divided overall into four groups based on the yearly survey areas. Therefore, there was no evidence for the different impact of vitamin A-enhanced transgenic soybean on the above-ground insects and spiders compared to non-GM soybean.

• Korean Journal of Environmental Biology
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• v.39 no.4
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• pp.445-451
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• 2021

In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

• Food Science and Industry
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• v.52 no.2
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• pp.140-152
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• 2019

"Contained use" means using LMOs during manufacturing processes within a facility, equipment or other physical structures to prevent the spread of LMOs to the external environment. LMOs and their substances not only have played an essential role in various industries in different ways, but also have made to spread concerns in Biosafety widely. In Korea, the LMO Act is to implement the Cartagena Protocol on Biosafety' since 1st Jan 2008. This is focused on Biosafety and Risk management for LMOs according to the precautionary principle.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.146.1-146
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• 2003

Transgenic Nicotiana benthmiana plants harboring and expressing coat protein (CP) gene of Zucchini green mottle mosaic virus (ZGMMV) were generated for both virus-resistant screening and complementation analysis of related viruses and environmental safety assessment (SA) of living modified organism (LMO) purposes. Transformation of leaf disc of N. benthamiana was performed using Agrobacterium-mediated method and the pZGCPPGA748 containing the ZGMMV CP and NPTII genes. Two kinds of transgenic homozygous groups, virus-resistant and -susceptible lines, were obtained by screening of challenging homologous virus for T1 generations. Complementation of CP-deficient related virus was analyzed using the susceptible line of ZGMMV. These two pathologically different lines can be useful for host-virus interactions and LMO environmental SA.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• v.2 no.2
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• pp.129-138
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• 2021

Living modified organisms (LMOs) are managed by seven government agencies according to their use in South Korea. The Ministry of Environment is responsible for LMOs used for environmental remediation. This study aimed to develop guidelines for assessing potential risks posed by transgenic plants used for remediation to insect ecosystems by investigating arthropod communities in sunflower fields. A total of 2,350 insects and spiders belonging to 134 species of 10 orders and 71 families were collected from sunflower fields over four growth stages ranging from anthesis to seed maturity. At the R3 phase of flower-bud formation, Chironomidae sp. of a decomposer insect guild presented the highest density, while Apis mellifera of a pollinator guild was the most abundant in the R5.8 phase of flowering. During the R7 seed-filling phase and the R9 phase of seed maturity, herbivorous Pochazia shantungensis predominated. During the R9 phase, richness and diversity indices of arthropod communities were distinctly lower whereas their dominance indices were significantly higher than those at other phases. In addition, the composition of arthropod communities was strongly correlated not only with the sampling date, but also with the sampling method depending on the growth stage of sunflowers. Our results suggest that appropriate sampling timing and methods should be considered in advance and that long-term field trials that cover a variety of environmental conditions should be carried out to evaluate potential risks to insect ecosystems.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• v.2 no.3
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• pp.210-218
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• 2021

Recently, pest-resistant living modified (LM) crops developed using RNA interference (RNAi) technology have been imported into South Korea. However, the potential adverse effects of unintentionally released RNAi-based LM crops on non-target species have not yet been reported. Coccinella septempunctata, which feeds on aphids, is an important natural enemy insect which can be exposed to the double-stranded RNA (dsRNA) produced by RNAi-based LM plants. To assess the risk of ingestion of Snf7 dsRNA by C. septempunctata, we first identified the species through morphological analysis of collected insects. A method for species identification at the gene level was developed using a specific C. septempunctata 12S rRNA. Furthermore, an experimental model was devised to assess the risk of Snf7 dsRNA ingestion in C. septempunctata. Snf7 dsRNA was mass-purified using an effective dsRNA synthesis method and its presence in C. septempunctata was confirmed after treatment with purified Snf7 dsRNA. Finally, the survival rate, development time, and dry weight of Snf7 dsRNA-treated C. septempunctata were compared with those of GFP and vATPase A dsRNA control treatments, and no risk was found. This study illustrates an effective Snf7 dsRNA synthesis method, as well as a high-concentration domestic insect risk assessment method which uses dsRNA to assess the risk of unintentional released of LM organisms against non-target species.

• Korean Journal of Environmental Biology
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• v.36 no.4
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• pp.488-497
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• 2018

Quantitative classification of insect fauna in transgenic rice in 2 LMO(Living genetically Modified Organism) isolated paddy fields (Gyeongsang National University, Gyeongbuk National University) and rice in 4 paddy fields(Duryang 1, Duryang 2, Hwagye 1, Hwagye 2), were evaluated for consecutive 5 years (2013 to 2017) in Sacheon, Gyeongnam province, and for 2 consecutive years (2015 to 2016) in Gunwi Gyeongbuk province. Sampling insect fauna were evaluated by black light trapping, sticky trapping, visual surveying, and sweeping methods in each target paddy field, respectively. A total of 37,941 individuals, of 464 species from 15 orders, were collected in Sacheon for 5 years. A total of 10,030 individuals, of 366 species from 13 orders, were collected in Gunwi for 2 years. Based on results of comparison between transgenic and non-transgenic paddy fields for 5 consecutive years, the similarity index between LMO paddy field and common paddy field is not different. Thus, this difference is not due to the environment, not the LMO.