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Genetically Engineered Mouse Models for Drug Development and Preclinical Trials

  • Lee, Ho
    • Biomolecules & Therapeutics
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    • v.22 no.4
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    • pp.267-274
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    • 2014
  • Drug development and preclinical trials are challenging processes and more than 80% to 90% of drug candidates fail to gain approval from the United States Food and Drug Administration. Predictive and efficient tools are required to discover high quality targets and increase the probability of success in the process of new drug development. One such solution to the challenges faced in the development of new drugs and combination therapies is the use of low-cost and experimentally manageable in vivo animal models. Since the 1980's, scientists have been able to genetically modify the mouse genome by removing or replacing a specific gene, which has improved the identification and validation of target genes of interest. Now genetically engineered mouse models (GEMMs) are widely used and have proved to be a powerful tool in drug discovery processes. This review particularly covers recent fascinating technologies for drug discovery and preclinical trials, targeted transgenesis and RNAi mouse, including application and combination of inducible system. Improvements in technologies and the development of new GEMMs are expected to guide future applications of these models to drug discovery and preclinical trials.

Mouse models for hepatitis B virus research

  • Hwang, Jeong-Ryul;Park, Sung-Gyoo
    • Laboraroty Animal Research
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    • v.34 no.3
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    • pp.85-91
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    • 2018
  • Hepatitis B virus (HBV) infection remains a major global health problem; indeed, there are 250 million carriers worldwide. The host range of HBV is narrow; therefore, few primates are susceptible to HBV infection. However, ethical constraints, high cost, and large size limit the use of primates as suitable animal models. Thus, in vivo testing of therapies that target HBV has been hampered by the lack of an appropriate in vivo research model. To address this, mouse model systems of HBV are being developed and several are used for studying HBV in vivo. In this review, we summarize the currently available mouse models, including HBV transgenic mice, hydrodynamic injection-mediated HBV replicon delivery systems, adeno-associated virus-mediated HBV replicon delivery systems, and human liver chimeric mouse models. These developed (or being developed) mouse model systems are promising and should be useful tools for studying HBV.

Comparative Study on Development of Mouse Embryos in Three Commercial Media and Hatching Rates of Mouse Embryos with/without Pronase (3개의 배양액내에서 생쥐배아의 발달과 Pronase로 처리한 생쥐배아 부화율의 비교 연구)

  • Lee, Jeong-Heon;Go, Hee-Jeong;Chae, Geu-Jeong;Lee, Ki-Suk;Kim, Jong-Duk
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.3
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    • pp.235-245
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    • 2001
  • Objectives: The purpose of this present study was to compare mouse embryo development in 3 commercial media and hatching competence of mouse embryo with or without enzymatic treatment. Methods: Collected 375 mouse embryos were divided into three groups, and then cultured in IVF-20 (G2), Medicult IVF (M3), P-1 (blastocyst M), respectively. Three day mouse morulae were cultured in G2 media treated with pronase. The results were analyzed using Chi-square test, and considered statistically significant when p<0.01. Results: The developmental rate of 2 cell mouse embryo after 72 hours was highest in IVF-20 (G2) among conventional 3 media. The hatching rate of mouse morulae was low when clultured in G2 media without pronase during 48 hours. However, it was higher when cultured in media treated with $1{\mu}g/ml$, $2.5{\mu}g/ml$, $5{\mu}g/ml$ pronase, respectively. Conclusions: Using good media and digestion of zona pellucida with enzymatic treatment improve development and hatching rate of embryo. Therefore, implantation and pregnancy rate could be improved.

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Obtainment and Characterization of Brain Tumor Cell Using Vasopressin-SV40 T Ag Transgenic Mouse

  • Kim, Sung-Hyun;Lee, Eun-Ju;Kim, Myoung-Li;Park, Jun-Hong;Cho, Kyoungin;Jung, Boo-Kyung;Kim, Hee-Chul;Hwnag, Sol-Ha;Lee, Hoon-Taek;Ryoo, Zae-Young
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • pp.105-105
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    • 2003
  • In previous reports, pVPSV.IGR2.1 transgenic mouse were described that brain tumor and lymphoma by reason of Vasopressin-SV40 T antigen. In this study, we produced pVPSV.IGR3.6 transgenic mouse that used pVPSV.IGR3.6 vector. Expression of transgene was vary different in transgenic mouse. We obtained 6 transgenic mouse line, moreover they had died at the age of 2-6 weeks without transmitting the transgene to their offspring, and had tumorigenesis on same location with pVPSV.IGR2.1 transgenic mouse. Only a founder mouse was investigated for expression of fusion gene. Here we extended this transgenic approach to the study of tumor progression. From the mouse, we confirmed brain tumor cell, after then cultured for investigate characterization. In this report, we demonstrate that reduction of survival rate in transgenic mouse fused vasopressin gene length, acquisition of brain tumor cell, composition with astrocyte cells and neuronal cells. Finally, cells had no change with increase of passage.

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Fabrication of fiber-optic evanescent wave immunosensor and its measuring characteristics (광섬유 소산파를 이용한 면역 센서 제조 및 그 특성)

  • Choi, Ki-Bong;Youn, Hee-Ju;Cha, Seung-Hee;Choi, Jung-Do
    • Journal of Sensor Science and Technology
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    • v.6 no.5
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    • pp.356-361
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    • 1997
  • Fiber-optic evanescent wave sensor was designed and fabricated to detect mouse immunoglobulin G(IgG) with decladed optical fiber on which anti-mouse IgG was immobilized. A sensitivity obtained by any direct or competitive method was lower than $1\;{\mu}g/m{\ell}$. Anti-mouse IgG was immobilized on 93.9% of core surface of optical fiber by simple adsorption method. The effect of postcoating using bovine serum albumin to remove non-specific binding was not observed. As the ratio of fluorescein to mouse IgG increased, the fluorescence signal increased, but that increase showed no linear relationship. Our fiber-optic sensor system could be used as immunosensor by measuring evanescent fluorescence in antigen-antibody reaction with good sensitivity below $1{\mu}g/m{\ell}$ level.

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Inflatable Mouse: Volume-adjustable Mouse with Air-pressure-sensitive Input and Haptic Feedback (부풀어지는 마우스: 기압센서를 이용한 입력과 햅틱 피드백을 갖는 부피가 변하는 마우스)

  • Kim, Seok-Tae;Lee, Bo-Ram;Kim, Hyun-Jung;Nam, Tek-Jin;Lee, Woo-Hun
    • 한국HCI학회:학술대회논문집
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    • pp.323-328
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    • 2008
  • Inflatable Mouse is a volume-adjustable user interface. It can be inflated up to the volume of a familiar mouse, but be deflated and stored flat in a PC card slot of a laptop computer when not in use. Inflatable Mouse functions just like a typical mouse; moreover, it provides new interaction techniques by sensing the air pressure in the balloon of the mouse. It also addresses some issues associated with pressure-sensing interactions such as the lack of bi-directional control and the lack of effective feedback. Moreover, it can be used as both a control tool and a display tool. In this paper, the design of an Inflatable Mouse prototype is described and potential application scenarios such as zooming in/out and fast scrolling using pressure control are explained. We also discuss the potential use of Inflatable Mouse as an emotional communication tool.

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Effect of Yanghyeuljeseuptang on immunological factors in spleen and draining lymph node(DLN) of atopic dermatitis induced NC/Nga mouse by dinitrochlorobenzene(DNCB) (양혈제습탕(凉血除濕湯)이 아토피 피부염 유발 NC/Nga mouse의 비장 및 DLN내 면역 관련 인자에 미치는 영향)

  • Park, Doo-Byoung;Han, Jae-Kyung;Kim, Yun-Hee
    • Journal of Haehwa Medicine
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    • v.16 no.2
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    • pp.251-265
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    • 2007
  • Yanhyeoljeseuptang(YHJST) is a traditional herbal medicine used for the treatment of dermatitis. The aim of this study was to confirm whether or not YHJST has a preventive effect on development of atopic dermatitis in dinitrochlorobenzene(DNCB)-applied Nc/Nga mouse. This study was undertaken to develop a reliable mouse model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. NC/Nga mouse were sensitized with $200\;{\mu\ell}$ of 1% 2,4-dinitrochlorobenzene(DNCB) (acetone : olive oil = 3 : 1 mixture) and challenged twice or three times with $150\;{\mu\ell}$ of 0.2% DNCB in a week for the following 4 weeks. YHJST was administered orally to Nc/Nga mouse for 8 weeks, which led to the remarkable suppression on the development of dermatitis, as determined by various immune factors related to pathogenesis of atopic dermatitis in splenocytes and DLN cells. In this study, YHJST selectively suppressed T ce11 (CD4+, CD3+/CD69+, CD4+/CD25+) activation, which may be essential for ratio of IL-4 versus INF-$\gamma$ produced in the splenic T cell culture supernatants was approximately 3-fold higher in the mouse treated with DNCB than their control mouse respectively. Immunologic studies showed down-regulated that the capacity of spleen T cells to produce IL-4, but IFN-$\gamma$ was up-regulated by means of oral intake of these YHJST. These results strongly suggest that YHJST is a promising candidate for treatment of human atopic dermatitis.

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Effect of Gupoongjeseuptang on immunological factors in spleen and draining lymph node(DLN) of atopic dermatitis induced NC/Nga mouse by dinitrochlorobenzene(DNCB) (구풍제습탕(驅風除濕湯)이 아토피 피부염 유발 NC/Nga mouse의 비장 및 DLN내 면역 관련 인자에 미치는 영향)

  • Yoon, Je-Eun;Han, Jae-Kyung;Kim, Yun-Hee
    • Journal of Haehwa Medicine
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    • v.16 no.2
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    • pp.267-280
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    • 2007
  • Gupoongjeseuptang(GPJST) is a traditional herbal medicine used for the treatment of dermatitis. The aim of this study was to confirm whether or not GPJST has a preventive effect on development of atopic dermatitis in dinitrochlorobenzene(DNCB)-applied Nc/Nga mouse. This study was undertaken to develop a reliable mouse model demonstrating similar immunologic phenomena as human atopic dermatitis characterized with predominance of type-2 immune response. NC/Nga mouse were sensitized with $200\;{\mu\ell}$ of 1% 2,4-dinitrochlorobenzene(DNCB) (acetone : olive oil = 3 : 1 mixture) and challenged twice or three times with $150\;{\mu\ell}$ of 0.2% DNCB in a week for the following 4 weeks. GPJST was administered orally to Nc/Nga mouse for 6 weeks, which led to the remarkable suppression on the development of dermatitis, as determined by various immune factors related to pathogenesis of atopic dermatitis in splenocytes and DLN cells. In this study, GPJST selectively suppressed T ce11 (CD4+, CD3+CD69+, CD4+CD25+) activation, which may be essential for ratio of IL-4 versus INF-$\gamma$ produced in the splenic T cell culture supernatants was approximately 3-fold higher in the mouse treated with DNCB than their control mouse respectively. Immunologic studies showed down-regulated that the capacity of spleen T cells to produce IL-4, but IFN-$\gamma$ was up-regulated by means of oral intake of these GPJST. These results strongly suggest that GPJST is a promising candidate for treatment of human atopic dermatitis.

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Characterization of Bovine NANOG5'-flanking Region during Differentiation of Mouse Embryonic Stem Cells

  • Jang, Hye-Jeong;Park, Hwan Hee;Tran, Thi Thuy Linh;Lee, Hak-Kyo;Song, Ki-Duk;Lee, Woon Kyu
    • Asian-Australasian Journal of Animal Sciences
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    • v.28 no.12
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    • pp.1721-1728
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    • 2015
  • Embryonic stem cells (ESCs) have been used as a powerful tool for research including gene manipulated animal models and the study of developmental gene regulation. Among the critical regulatory factors that maintain the pluripotency and self-renewal of undifferentiated ESCs, NANOG plays a very important role. Nevertheless, because pluripotency maintaining factors and specific markers for livestock ESCs have not yet been probed, few studies of the NANOG gene from domestic animals including bovine have been reported. Therefore, we chose mouse ESCs in order to understand and compare NANOG expression between bovine, human, and mouse during ESCs differentiation. We cloned a 600 bp (-420/+181) bovine NANOG 5'-flanking region, and tagged it with humanized recombinant green fluorescent protein (hrGFP) as a tracing reporter. Very high GFP expression for bovine NANOG promoter was observed in the mouse ESC line. GFP expression was monitored upon ESC differentiation and was gradually reduced along with differentiation toward neurons and adipocyte cells. Activity of bovine NANOG (-420/+181) promoter was compared with already known mouse and human NANOG promoters in mouse ESC and they were likely to show a similar pattern of regulation. In conclusion, bovine NANOG 5-flanking region functions in mouse ES cells and has characteristics similar to those of mouse and human. These results suggest that bovine gene function studied in mouse ES cells should be evaluated and extrapolated for application to characterization of bovine ES cells.

An Arachidonic Acid Metabolizing Enzyme, 8S-Lipoxygenase, in Mouse Skin Carcinogenesis

  • Kim Eun-Jung
    • Nutritional Sciences
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    • v.9 no.3
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    • pp.212-226
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    • 2006
  • The involvement of arachidonic acid (AA) metabolizing enzyme, lipoxygenase (LOX), in the development of particular tumors in humans has gradually been acknowledged and LOX has emerged as a novel target to prevent or treat human cancers. In the mouse skin carcinogenesis model, which provides an excellent model to study multistage nature of human cancer development, many studies have shown that some of the LOXs are constitutively upregulated in their expression. Moreover, application of LOX inhibitors effectively reduced tumor burdens, which implicates the involvement of LOX in mouse skin tumor development as well. 8S-LOX is a recently cloned LOX, which is specifically expressed in mouse skin after 12-O-tetradecanoyl-phorbol-13-acetate (TPA) treatment but not in normal skin. Unlike other members of the LOX 'family' expressed in mouse skin, this TPA-induced expression of 8S-LOX is prominent only in the skin of the TPA tumor promotion-sensitive strains of mice (SENCAR, CD-1, and NMRI) but not in the promotion-resistant C57BL/6J mice. This is a very unique phenomenon among strains of mice. Constitutive upregulation of 8S-LOX was also found in early stage papillomas and the expression was gradually reduced as the tumors became malignant. Based on these observations, it has been thought that 8S-LOX is involved in TPA-induced tumor promotion as well as in tumor conversion from papillomas to carcinomas. In accordance with this hypothesis, several studies have suggested possible roles of 8S-hydroxyeicosatetraenoic acid (HETE), an AA metabolite of 8S-LOX, in mouse skin tumor development. A clastogenic activity of 8S-HETE was demonstrated in primary keratinocytes and a close correlation between the levels of etheno-DNA adducts and 8S-HETE during skin carcinogenesis was also reported. On the other hand, it has been reported that 8S-LOX protein expression is restricted to a differentiated keratinocyte compartment Moreover, reported findings on the ability of 8S-HETE to cause keratinocyte differentiation appear to be contrary to the procarcinogenic features of the 8S-LOX expression, presenting a question as to the role of 8S-LOX during mouse skin carcinogenesis. In this review, molecular and biological features of 8S-LOX as well as current views on the functional role of 8S-LOX/8S-HETE during mouse skin carcinogenesis are presented.