• Molecules and Cells
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• 제22권3호
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• pp.275-284
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• 2006

The molecular components that generate and maintain circadian rhythms of physiology and behavior in mammals are present both in the brain (suprachiasmatic nucleus; SCN) and in peripheral tissues. Examination of mice with targeted disruptions of either mPer1 or mPer2 has shown that these two genes have key roles in the SCN circadian clock. Here we show that loss of the clock gene mPer2 affects forced locomotor performance in mice without altering muscle contractility. A proteomic analysis revealed that the anterior tibialis muscles of the mPer2 knockout mice had higher levels of glycolytic enzymes such as triose phosphate isomerase and enolase than those of either the wild type or mPer1 knockout mice. In addition, the level of expression of HSP90 in the mPer2 mutant mice was also significantly higher than in wildtype mice. These results suggest that the reduced locomotor endurance of the mPer2 knockout mice reflects a greater dependence on anaerobic metabolism under stress conditions, and that the two canonical clock genes, mPer1 and mPer2, play distinct roles in the physiology of skeletal muscle.

• 한국발생생물학회지:발생과생식
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• 제28권2호
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• pp.29-36
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• 2024

Cellular prion protein (PrP^C) encoded at Prnp gene is well-known to form a misfolded isoform, termed scrapie PrP (PrP^SC) that cause transmissible degenerative diseases in central nervous system. The physiological role of PrP^C has been proposed by many studies, showing that PrP^C interacts with various intracellular, membrane, and extracellular molecules including mitochondrial inner membrane as a scaffold. PrP^C is expressed in most cell types including reproductive organs. Numerous studies using PrP^C knockout rodent models found no obvious phenotypic changes, in particular the clear phenotypes in development and reproduction have not demonstrated in these knockout models. However, various roles of PrP^C have been evaluated at the cellular levels. In this review, we summarized the known roles of PrP^C in various cell types and tissues with a special emphasis on those involved in reproduction.

• Interdisciplinary Bio Central
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• 제2권4호
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• pp.15.1-15.5
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• 2010

RIKEN BioResource Center (BRC) has collected, preserved, conducted quality control of, and distributed mouse resources since 2002 as the core facility of the National BioResource Project by the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan. Our mouse resources include over 5,000 strains such as humanized disease models, fluorescent reporters, and knockout mice. We have developed novel mouse strains such as tissue-specific Cre-drivers and optogenetic strains that are in high demand by the research community. We have removed all our specified pathogens from the deposited mice and used our quality control tests to examine their genetic modifications and backgrounds. RIKEN BRC is a founding member of the Federation of International Mouse Resources and the Asian Mouse Mutagenesis and Resource Association, and provides mouse resources to the one-stop International Mouse Strain Resource database. RIKEN BRC also participates in the International Gene Trap Consortium, having registered 713 gene-trap clones and their sequences in a public library, and is an advisory member of the CREATE (Coordination of resources for conditional expression of mutated mouse alleles) consortium which represents major European and international mouse database holders for the integration and dissemination of Cre-driver strains. RIKEN BRC provides training courses in the use of advanced technologies for the quality control and cryopreservation of mouse strains to promote the effective use of mouse resources worldwide.

• Mycobiology
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• 제47권2호
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• pp.207-216
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• 2019

Talaromyces marneffei is the only dimorphic species in its genus and causes a fatal systemic mycosis named talaromycosis. Our previous study indicated that knockdown of AcuD gene (encodes isocitrate lyase of glyoxylate bypass) of T. marneffei by RNA interference approach attenuated the virulence of T. marneffei, while the virulence of the AcuD knockout strains was not studied. In this study, T. marneffei-zebrafish infection model was successfully established through hindbrain microinjection with different amounts of T. marneffei yeast cells. After co-incubated at $28^{\circ}C$, the increasing T. marneffei inoculum doses result in greater larval mortality; and hyphae generation might be one virulence factor involved in T. marneffei-zebrafish infection. Moreover, the results demonstrated that the virulence of the ${\Delta}AcuD$ was significantly attenuated in this Zebrafish infection model.

• Asian-Australasian Journal of Animal Sciences
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• 제30권8호
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• pp.1183-1189
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• 2017

Objective: Owing to the public availability of complete genome sequences, including avian species, massive bioinformatics analyses may be conducted for computational gene prediction and the identification of gene regulatory networks through various informatics tools. However, to evaluate the biofunctional activity of a predicted target gene, in vivo and in vitro functional genomic analyses should be a prerequisite. Methods: Due to a lack of quail genomic sequence information, we first identified the partial genomic structure and sequences of the quail SH3 domain containing ring finger 2 (SH3RF2) gene. Subsequently, SH3RF2 was knocked out using clustered regularly interspaced short palindromic repeat/Cas9 technology and single cell-derived SH3RF2 mutant sublines were established to study the biofunctional activity of SH3RF2 in quail myoblast (QM7) cells during muscle differentiation. Results: Through a T7 endonuclease I assay and genotyping analysis, we established an SH3RF2 knockout (KO) QM7#4 subline with 61 and 155 nucleotide deletion mutations in SH3RF2. After the induction of myotube differentiation, the expression profiles were analyzed and compared between regular QM7 and SH3RF2 KO QM7#4 cells by global RNA sequencing and bioinformatics analysis. Conclusion: We did not detect any statistically significant role of SH3RF2 during myotube differentiation in QM7 myoblast cells. However, additional experiments are necessary to examine the biofunctional activity of SH3RF2 in cell proliferation and muscle growth.

• 한국독성학회:학술대회논문집
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• 한국독성학회 1998년도 국제심포지움 및 추계학술대회
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• pp.8-13
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• 1998

Polyoma Virus Enhancer core Binding Protein (PEBP2)는 유전자의 전사를 조절하는 hetero-dimeric transcription factor로서 $\alpha$와 $\beta$ subunit으로 구성되어 있다. $\alpha$ subunit을 coding 하는 유전자중 하나인 PEBP2aB는 급성백혈병과 관련되어있는 t(8;21) 또는 t(12;21)에 의하여 변형됨으로서 백혈병 발병의 원인이 되고 있다 (Miyoshi et al., 1993; .Romana et al., 1995). $\beta$ subunit을 coding 하는 PEBP2$\beta$도 inv(16)에 의하여 변형됨으로서 백혈병을 유도하는 주요 원인이 되고 있다 (Liu et al., 1993). 이 유전자들의 생물학적 활성을 밝히기 위한 연구가 gene targeting에 의한 knockout mouse 생산 방법으로 수행되었다. 그 결과 PEBP2$\alpha$B와 PEBP2$\beta$ 유전자가 definitive hematopoiesis에 있어서 결정적으로 중요한 역할을 하고 있음이 관찰되었다 (Okuda et al., 1996, Wang et al., 1996a, 1996b), 이는 이들 유전자가 bematopoietic master switch 유전자임을 밝힌 중요한 결과로서 이로부터 혈액학 연구 분야의 새로운 장이 열리게 되었다. 또한 이러한 연구 결과들은 PEBP2 family에 속하는 다른 유전자의 생물학적 활성의 연구를 촉진하는 계기가 되었다. 최근 PEBP2$\alpha$A 유전자가 결손된 마우스가 생산되었는데 이 유전자의 경우에는 모든 종류의 뼈의 생성이 완전히 결손됨이 관찰되었다 (Komori et al., 1997). 이는 PEBP2$\alpha$A 유전자가 뼈의 생성을 지배하는 master switch 유전자임을 보여주는 중요한 관찰로서 bone biologist 들의 큰 관심을 모으고 있다. 본 연구팀은 PEBP2 family 유전자 중 유일하게 아직 생물학적 활성이 규명되지 않은 PEBP2$\alpha$C 유전자의 활성을 knockout 마우스를 생산하는 방법에 의하여 분석하였으며 소화기관의 형성에 중요한 역할을 하고 있음을 확인하였다.

• 대한방사성의약품학회지
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• 제4권1호
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• pp.26-31
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• 2018

Macrophages play a key role in atherosclerotic plaque formation, but their participation has been discerned largely via ex vivo analyses of atherosclerotic lesions. Therefore, we aimed to identify atherosclerosis on noninvasive in vivo imaging using reporter gene system. This study demonstrated that recruitment of macrophages could be detected in atherosclerotic plaques of Apolipoprotein E knockout (ApoE-/-) mice with a sodium iodide symporter (NIS) gene imaging system using $^{99m}Tc-SPECT$. This novel approach to tracking macrophages to atherosclerotic plaques in vivo could have applications in studies of arteriosclerotic vascular disease.

• Animal Bioscience
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• 제36권2_spc호
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• pp.333-338
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• 2023

Genetic modification enables modification of target genes or genome structure in livestock and experimental animals. These technologies have not only advanced bioscience but also improved agricultural productivity. To introduce a foreign transgene, the piggyBac transposon element/transposase system could be used for production of transgenic animals and specific target protein-expressing animal cells. In addition, the clustered regularly interspaced short palindromic repeat-CRISPR associated protein 9 (CRISPR-Cas9) system have been utilized to generate chickens with knockout of G0/G1 switch gene 2 (G0S2) and myostatin, which are related to lipid deposition and muscle growth, respectively. These experimental chickens could be the invaluable genetic resources to investigate the regulatory pathways and mechanisms of improvement of economic traits such as fat quantity and growth. The gene-edited animals could also be applicable to the livestock industry.

• 원예과학기술지
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• 제30권6호
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• pp.734-742
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• 2012

본 연구는 배추의 유전자 기능분석을 위한 RNAi 유전자 침묵 기법과 T-DNA 삽입 기법을 비교하기 위해 수행하였다. 두 종류의 형질전환 계통이 이용되었으며 BrSAMS-knockout(KO) 계통은 T-DNA 삽입으로 한 개의 Brassica rapa S-adenosylmethionine synthetase(BrSAMS) 유전자가 기능을 상실한 계통이었으며 BrSAMS-knockdown(KD) 계통은 RNAi 방법을 통해 BrSAMS 유전자들의 발현이 억제된 계통이었다. KO 계통과 KD 계통의 microarray 분석 결과에서는 SAMS 유전자와 관련된 sterol, 자당, homogalacturonan 생합성 및 glutaredoxin-related protein, serine/threonine protein kinase, 그리고 gibberellin-responsive protein 유전자들의 발현 수준이 뚜렷한 차이를 보여 주었다. 그러나 KO 계통의 유전자 발현 양상은 하나의 BrSAMS 유전자가 기능을 상실하였음에도 불구하고 대조 계통과 비교하여 RNAi기법을 적용한 KD 계통에 비해 큰 차이를 보여주지 못했다. 또한 직접적으로 SAMS 유전자와 관련된 폴리아민과 에틸렌 합성 유전자들의 발현 변화도 KD 계통에서 더 잘 나타났다. 본 연구에서 microarray 결과를 이용한 KO 계통의 BrSAMS 기능분석은 배추과식물의 게놈 triplication 발생으로 인하여 다수로 존재하는 SAMS 유전자들 때문에 명확한 결론을 얻을 수 없었다. 결론적으로 배추와 같은 배수체 작물의 유전자 기능 분석은 RNAi silencing에 의한 유전자 knock-down 기법이 T-DNA 삽입에 의한 knock-out 기법보다 더욱 효율적인 것으로 나타났다.

• 한국환경성돌연변이발암원학회:학술대회논문집
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• 한국환경성돌연변이발암원학회 2002년도 Current Trends in Toxicological Sciences
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• pp.36-42
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• 2002

Diesel exhaust (DE) has been recognized as a noxious mutagen and/or carcinogen, because its components can form DNA adducts. Mechanisms governing the susceptibility to DE and the efficiency of such DNA adduct formation require clarification. The transcription factor Nrf2 is essential for inducible and/or constitutive expression of a group of detoxification and antioxidant enzymes, and we hypothesized that the nrf2 gene knockout mouse might serve as an excellent model system for analyzing DE toxicity. To address this hypothesis, lungs from nrf2(-/-) and nrf2(+/-) mice were examined for the production of xenobiotic-DNA adducts after exposure to DE (3 $mg/m^{3}$ suspended particulate matter) for 4 weeks. Whereas the relative adduct levels (RAL) were significantly increased in the lungs of both nrf2(+/-) and nrf2(-/-) mice upon exposure to DE, the increase of RAL in the lungs from nrf2(-/-) mice exposed to DE were approximately 2.3-fold higher than that of nrf2(+/-) mite exposed to DE. In contrail, cytochrome P4501Al mRNA levels in the nrf2(-/-)mouse lungs were similar to those in the nrf2(+/-) mouse lungs even after exposure to DE, suggesting that suppressed activity of phase II drug-metabolizing enzymes is important in giving ise to the increased level of DNA adducts in the Nrf2-null mutant mouse subjected to DE. Importantly, severe hyperplasia and accumulation of the oxidative DNA adduct 8-hydroxydeoxyguanosine were observed in the bronchial epidermis of nrf(-/-) mite following DE exposure. These results demonstrate the increased susceptibility of the nrf2 germ line mutant mouse to DE exposure and indicate the nrf2 gene knockout mouse nay represent a valuable model for the assessment of respiratory DE toxicity.