• Title/Summary/Keyword: Gene Targeted

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Improvement of Seedling Establishment in Wet Direct Seeding of Rice using the Anaerobic Germination Tolerance Gene Derived from Weedy Photoblastic Rice (잡초벼 PBR 혐기발아 내성 유전자 활용 벼 담수직파 초기 입모 개선)

  • Jeong, Jong-Min;Mo, Youngjun;Baek, Man-Kee;Kim, Woo-Jae;Cho, Young-Chan;Ha, Su-Kyung;Kim, Jinhee;Jeung, Ji-Ung;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.161-171
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    • 2020
  • Direct seeding is one of the rice seedling establishment methods that is increasingly being practiced by farmers to save labor and reduce costs. However, this method often causes poor germination under flooding conditions after sowing. In this study, we developed japonica elite lines with quantitative trait loci (QTL) associated with anaerobic germination (AG) tolerance to overcome poor germination and seedling establishment in wet direct seeding. The QTL introgression lines were developed from a cross between weedy photoblastic rice as the AG donor and the Nampyeong variety via phenotypic and genotypic selection. Compared to Nampyeong, the survival rates of the selected lines were improved by approximately 50% and 240% under field and greenhouse conditions, respectively. To improve selection efficiency by marker assisted selection, the QTL markers associated with AG tolerance were converted to cleaved amplified polymorphic sequence markers designed based on next-generation sequence analysis. These lines retained similar agronomic traits and yield potential to the parent, Nampyeong. Among these lines, we selected the most promising line, which exhibited high survival rate and good agricultural traits under flooding conditions and named the line as Jeonju643. This line will contribute to breeding programs aiming to develop rice cultivars adapted to wet direct seeding. This study demonstrates the successful application of marker-assisted selection to targeted introgression of anaerobic genes into a premium quality japonica rice variety.

An International Collaborative Program To Discover New Drugs from Tropical Biodiversity of Vietnam and Laos

  • Soejarto, Djaja D.;Pezzuto, John M.;Fong, Harry H.S.;Tan, Ghee Teng;Zhang, Hong Jie;Tamez, Pamela;Aydogmus, Zeynep;Chien, Nguyen Quyet;Franzblau, Scott G.;Gyllenhaal, Charlotte;Regalado, Jacinto C.;Hung, Nguyen Van;Hoang, Vu Dinh;Hiep, Nguyen Tien;Xuan, Le Thi;Hai, Nong Van;Cuong, Nguyen Manh;Bich, Truong Quang;Loc, Phan Ke;Vu, Bui Minh;Southavong, Boun Hoong
    • Natural Product Sciences
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    • v.8 no.1
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    • pp.1-15
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    • 2002
  • An International Cooperative Biodiversity Group (ICBG) program based at the University of Illinois at Chicago initiated its activities in 1998, with the following specific objectives: (a) inventory and conservation of of plants of Cuc Phuong National Park in Vietnam and of medicinal plants of Laos; (b) drug discovery (and development) based on plants of Vietnam and Laos; and (c) economic development of communities participating in the ICBG project both in Vietnam and Laos. Member-institutions and an industrial partner of this ICBG are bound by a Memorandum of Agreement that recognizes property and intellectual property rights, prior informed consent for access to genetic resources and to indigenous knowledge, the sharing of benefits that may arise from the drug discovery effort, and the provision of short-term and long-term benefits to host country institutions and communities. The drug discovery effort is targeted to the search for agents for therapies against malaria (antimalarial assay of plant extracts, using Plasmodium falciparum clones), AIDS (anti-HIV-l activity using HOG.R5 reporter cell line (through transactivation of the green fluorescent protein/GFP gene), cancer (screening of plant extracts in 6 human tumor cell lines - KB, Col-2, LU-l, LNCaP, HUVEC, hTert-RPEl), tuberculosis (screening of extracts in the microplate Alamar Blue assay against Mycobacterium tuberculosis $H_{37}Ra\;and\;H_{37}Rv),$ all performed at UIC, and CNS-related diseases (with special focus on Alzheimer's disease, pain and rheumatoid arthritis, and asthma), peformed at Glaxo Smith Kline (UK). Source plants were selected based on two approaches: biodiversity-based (plants of Cuc Phuong National Park) and ethnobotany-based (medicinal plants of Cuc Phuong National Park in Vietnam and medicinal plants of Laos). At mc, as of July, 2001, active leads had been identified in the anti-HIV, anticancer, antimalarial, and anti- TB assay, after the screening of more than 800 extracts. At least 25 biologically active compounds have been isolated, 13 of which are new with anti-HIV activity, and 3 also new with antimalarial activity. At GSK of 21 plant samples with a history of use to treat CNS-related diseases tested to date, a number showed activity against one or more of the CNS assay targets used, but no new compounds have been isolated. The results of the drug discovery effort to date indicate that tropical plant diversity of Vietnam and Laos unquestionably harbors biologically active chemical entities, which, through further research, may eventually yield candidates for drug development. Although the substantial monetary benefit of the drug discovery process (royalties) is a long way off, the UIC ICBG program provides direct and real-term benefits to host country institutions and communities.

Human Parechovirus as an Important Cause of Central Nervous System Infection in Childhood (소아청소년기 중추신경 감염의 주요 원인으로서 Human Parechovirus의 의의)

  • Jung, Hyun Joo;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.23 no.3
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    • pp.165-171
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    • 2016
  • Purpose: Human parechovirus (HPeV) is an increasingly recognized pathogenic cause of central nervous system (CNS) infection in neonates. However, HPeV infections have not been studied in older children. This study determined the prevalence and clinical features of HPeV CNS infection in children in Korea. Methods: Reverse transcription polymerase chain reaction assays were performed using HPeV-specific, 5' untranslated, region-targeted primers to detect HPeV in cerebrospinal fluid (CSF) samples from children presenting with fever or neurologic symptoms from January 1, 2013, to July 31, 2014. HPeV genotyping was performed by sequencing the viral protein 3/1 region. Clinical and laboratory data were retrospectively abstracted from medical records and compared with those of enterovirus (EV)-positive patients from the same period. Results: Of 102 CSF samples, six (5.9%) were positive for HPeV; two of 21 EV-positive samples were co-infected with HPeV. All samples were genotype HPeV3. Two HPeV-positive patients were <3 months of age and four others were over 1 year old. While HPeV-positive infants under 1 year of age presented with sepsis-like illness without definite neurologic abnormalities, HPeV-positive children over 1 year of age presented with fever and neurologic symptoms such as seizures, loss of consciousness, and gait disturbance. The CSF findings of HPeV-positive patients were mostly within the normal range, whereas most (73.7%) EV-positive patients had pleocytosis. Conclusions: Although HPeV is typically associated with disease in young infants, the results of this study suggest that HPeV is an emerging pathogen of CNS infection with neurologic symptoms in older childhood.

Development and Validation of Real-time PCR to Determine Branchiostegus japonicus and B. albus Species Based on Mitochondrial DNA (Real-time PCR 분석법을 이용한 옥돔과 옥두어의 종 판별법 개발)

  • Chung, In Young;Seo, Yong Bae;Yang, Ji-Young;Kim, Gun-Do
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1331-1339
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    • 2017
  • DNA barcoding is the identification of a species based on the DNA sequence of a fragment of the cytochrome C oxidase subunit I (COI) gene in the mitochondrial genome. It is widely applied to assist with the sustainable development of fishery-product resources and the protection of fish biodiversity. This study attempted to verify horse-head fish (Branchiostegus japonicus) and fake horse-head fish (Branchiostegus albus) species, which are commonly consumed in Korea. For the validation of the two species, a real-time PCR method was developed based on the species' mitochondrial DNA genome. Inter-species variations in mitochondrial DNA were observed in a bioinformatics analysis of the mitochondrial genomic DNA sequences of the two species. Some highly conserved regions and a few other regions were identified in the mitochondrial COI of the species. In order to test whether variations in the sequences were definitive, primers that targeted the varied regions of COI were designed and applied to amplify the DNA using the real-time PCR system. Threshold-cycle (Ct) range results confirmed that the Ct ranges of the real-time PCR were identical to the expected species of origin. Efficiency, specificity and cross-reactivity assays showed statistically significant differences between the average Ct of B. japonicus DNA ($21.85{\pm}3.599$) and the average Ct of B. albus DNA ($33.49{\pm}1.183$) for confirming B. japonicus. The assays also showed statistically significant differences between the average Ct of B. albus DNA ($22.49{\pm}0.908$) and the average Ct of B. japonicus DNA ($33.93{\pm}0.479$) for confirming B. albus. The methodology was validated by using ten commercial samples. The genomic DNA-based molecular technique that used the real-time PCR was a reliable method for the taxonomic classification of animal tissues.

Alteration of MicroRNAs Targeted Integrins by PD-MSCs Transplantation Is Involved in Hepatic Regeneration in a Rat Model with BDL (담관결찰 쥐 모델에서 태반유래중간엽줄기세포 이식에 의한 miRNA 표적 인테그린 변화의 간재생 효과)

  • Park, Sohae
    • Journal of Life Science
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    • v.31 no.8
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    • pp.710-718
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    • 2021
  • Placenta-derived mesenchymal stem cells (PD-MSCs) are promising candidates for cell-based therapy in regenerative medicine. The migration and homing potential of PD-MSCs to injured sites is a critical property of MSC engraftment. MicroRNAs (miRNAs) have recently been shown to regulate the critical functions of MSCs, such as proliferation, survival, and migration. The objective of the present study was to identify the miRNA and target genes involved in PD-MSCs homing in a bile duct ligation (BDL) rat model. We selected candidate miRNAs targeting genes for PD-MSCs homing based on microarray analysis. PD-MSC engraftment in BDL-injured rat liver was identified by immunofluorescence assay and human-specific Alu gene expression by quantitative real-time polymerase chain reaction (qRT-PCR) one week after transplantation. Compared with migrated naïve PD-MSCs under hypoxic and normoxic conditions (Hyp/Nor), the transplanted group with PD-MSCs (Tx) showed distinct differences in miRNA expressions in BDL-injured rat liver. We also validated the miRNAs and their target genes for PD-MSCs homing. The expressions of integrin α4 (ITGA4) and integrin α5 (ITGA5) target genes for miR-199a-5p and miR-148a-3p were significantly upregulated in the Tx group (p<0.05). In addition, integrin β1 (ITGB1) and integrin β8 (ITGB8) were upregulated by suppressing miR-183-5p and miR-145-5p, respectively. These results demonstrated that PD-MSCs regulate miRNA expression related to the integrin family for their homing effects on the BDL-injured rat liver. The findings further suggest that miRNA-mediated regulation of the integrin family contributes to the therapeutic efficacy of PD-MSCs in the rat hepatic fibrosis model by BDL.

Enhancement of Protein Aggregate Clearance in Huntington's Disease Model viaCRISPR/dCas9 Activation of NAGK and Reln Genes (CRISPR/dCas9을 통한 NAGK 및 Reln 유전자 활성화에 의한 헌팅턴병 모델에서 단백질 응집체 제거 촉진)

  • Diyah Fatimah Oktaviani;Raju Dash;Sarmin Ummey Habiba;Ho Jin Choi;Yeasmin Akter Munni;Dae-Hyun Seog;Maria Dyah Nur Meinita;Il Soo Moon
    • Journal of Life Science
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    • v.34 no.9
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    • pp.609-619
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    • 2024
  • Neurodegenerative diseases are marked by the accumulation of toxic misfolded proteins in neurons. Therefore, strategies for the effective prevention and clearance of aggregates are crucial for therapeutic interventions. Cytoplasmic dynein plays a crucial role in the clearance of aggregates by transporting them to the cell center, where lysosomes are enriched and the aggregates undergo extensive autophagic degradation. Previously, we reported evidence for the activation of dynein by N-acetylglucosamine kinase (NAGK) and Reln. In the present study, we explored the effects of NAGK and Reln upregulation on the clearance of aggregates. To upregulate NAGK and Reln genes in HEK293T cells (a human embryonic kidney cell line), CRISPR/dCas9 activation systems (CASs) were used with specific plasmids encoding target-specific 20 nt guide RNA. The effects of this genetic modulation were analyzed in Huntington's disease cellular models, including HEK293T cells and primary mouse cortical cells, where external mutant huntingtin (mHtt, Q74) aggregates were induced. The results showed that the CAS activation of NAGK or Reln, or their combination, significantly reduced the proportion of cells with Q74 aggregates (aggresomes). This effect was reversed by Ciliobrevin D (a dynein inhibitor) and chloroquine (an autophagy inhibitor), indicating the role of dynein-mediated autophagy in aggregate clearance. These findings provide the basis for therapeutic strategies aimed at enhancing neuronal health through targeted gene activation.