• Title/Summary/Keyword: transgenic mice

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Human Erythropoietin Induces Lung Failure and Erythrocytosis in Transgenic Mice

  • Kim, Myoung Ok;Kim, Sung Hyun;Shin, Mi Jung;Lee, Dong Beom;Kim, Tae Won;Kim, Kil Soo;Ha, Ji Hong;Lee, Sanggyu;Park, Yong Bok;Kim, Sun Jung;Ryoo, Zae Young
    • Molecules and Cells
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    • v.23 no.1
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    • pp.17-22
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    • 2007
  • We have expressed human erythropoietin (EPO) in transgenic mice using a recombinant EPO cDNA combined with a partial TPO construct. The gene was microinjected using standard techniques and five mice were detected as transgenic by PCR and further used as founders. The life span of the transgenic founders was much shorter than that of their normal littermates. Most of the tissues of the transgenic founders contained human EPO transcripts as judged by RT-PCR. Especially high expression levels were seen in the liver and lung. EPO protein levels in serum were examined by ELISA and ranged from 266-414 mIU/ml. The number of red blood cell, white blood cell and hemoglobin in the hEPO transgenic mice was higher than in normal mice. These results indicate that overexpression of hEPO is deleterious and can provoke lung failure and erythrocytosis.

Expression of Antisense Mouse Obese Gene in Transgenic Mice (형질전환 생쥐에서 Antisense 비만유전자의 발현)

  • Kwon, B.S.;Hong, K.H.;Jahng, J.W.;Lee, H.T.;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.419-428
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    • 2000
  • Leptin, the product of obese (ob) gene, is an adipocyte-derived satiety factor that plays a major role in the regulation of food intake, energy homeostasis, body weight, reproductive physiology and neuropeptide secretion. The present study was designed to generate transgenic mice expressing antisense mouse ob (mob) gene. Total RNA was extracted from the adipose tissues of mouse, then reverse transcription was performed. The 303 and 635 bp fragments of anti I and II cDNAs were amplified from mob cDNAs by PCR. The two mob cDNAs were reversely ligated into between adipose tissue specific aP2 promote and SV40 poly(A) site. Transgenic mice carrying two different kinds of antisense mob transgenes were generated by DNA microinjection into pronucleus. Total 14 transgenic mice were born, and the 4 and 5 founder lines of the transgenic mice with anti I and II transgenes were respectively established. Antisense mRNA expression was detected in transgenic F$_1$ mice by RT-PCR analysis. This result suggests that the transgenic mice expressing antisense mob mRNA may be useful as an animal disease model to be obesity caused by decreased amount of leptin secretion.

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IL-4 Suppresses UVB-induced Apoptosis in Skin

  • Hwang, Ha-Young;Choi, Soo-Young;Kim, Tae-Yoon
    • BMB Reports
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    • v.40 no.1
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    • pp.36-43
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    • 2007
  • In this study, cutaneous role of IL-4 in UVB-induced apoptosis was investigated using transgenic mice with skin-specific expression of IL-4 (IL-4 Tg mice). The transgenic mice did not show any gross clinical abnormalities. However, epidermis was thickened and increased MHC class II positive cells were detected as well as enhanced expression of inflammatory cytokines such as IL-1 and TNF-$\alpha$ in skin. In addition, histological analysis revealed increased infiltration of lymphocytes, acanthosis, hyperkeratosis, and parakeratosis in skin of IL-4 Tg mice. The physiological effect of IL-4 overexpression in skin against environmental stimulus such as UVB was investigated by irradiating wild-type and IL-4 Tg mice with UVB followed by evaluation of apoptosis. The result demonstrated suppressed apoptosis in epidermis of IL-4 Tg mice compared with wild-type mice. To further assess anti-apoptotic function of IL-4 in keratinocytes, stable cell clones were made where IL-4 was constitutively overexpressed and examined for UVB-induced apoptosis. The results showed that apoptosis was remarkably decreased in IL-4 over-expressing cell clones compared with that in mock transfected cells. Collectively, data presented here shows that IL-4 has an inhibitory effect against UVB-induced apoptosis in keratinocytes, suggesting that IL-4 may be an important regulator in cutaneous immunity against UVB.

A Correspondence between Aging-related Reduction of Neprilysin and Elevation of Aβ-42 or γ-Secretase Activity in Transgenic Mice Expressing NSE-controlled APPsw or Human Mutant Presenilin-2

  • Lim Hwa-J.;Kim Yong-K.;Sheen Yhun-Y.
    • Biomolecules & Therapeutics
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    • v.14 no.2
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    • pp.106-109
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    • 2006
  • Neprilysin (Nep) is known to be important to degrade $A{\beta}$ derived from amyloid precursor protein (APP) by cleavage with $\beta-and\;\gamma$-secretases. In order to determine whether a correspondence between $A{\beta}-42/{\gamma}-secretase$ activity and Nep levels exists in postnatal aging of transgenic mice expressing either neuron-specific enolase (NSE)-controlled human mutant presenilin-2 (hPS2m) or APPsw alone, the levels of Nep expression and $A{\beta}-42/{\gamma}-secretase$ activity were examined age of 5, 12, and 20 months, respectively. The levels of Nep expression in both types of transgenic brains were decreased relative to those of control mice in a aging-related manner, while the level of $A{\beta}-42/{\gamma}-secretase$ activity was reversibly increased. Thus, changes in $A{\beta}-42$ may all reflect variation in amounts of Nep enzyme.

Co-expression of Human Proteins (IL-10, TPO and/or Lactoferrin) into Milk of Cross-Breed Transgenic Mouse

  • Zheng, Zhen-Yu;Lee, Hyo-Sang;Oh, Keon-Bong;Koo, Deog-Bon;Han, Yong-Mahn;Lee, Kyung-Kwang
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.45-49
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    • 2008
  • We have previously produced transgenic (TG) mice expressing the human lactoferrin (hLF), interleukin-10 (hIL-10), and thrombopoietin (hTPO) proteins in the milk. In this study, we examined whether simple crossbreeding between two kids of a single transgenic mouse can produce double transgenics co-expressing two human proteins.. The hLF male, and the hIL-10 male were crossbred with the hIL-10 and hTPO females, and the hTPO female, respectively. PCR analysis for genotyping showed 32%, 23% and 24% double transgenic rates for hLF/hIL-10, hLF/hTPO, and hIL-10/hTPO transgenes, respectively. We analyzed the expression levels of the human proteins from double transgenic mice and compared those with their single transgenic siblings. All double transgenic co-expressed two human proteins at comparable levels to singles', unless hTPO was not co-expressed: for hLF, 1.1 mg/ml in hLF/hIL-10, whereas 0.5 mg/ml in hLF/hTPO; for hIL-10, 4.1 mg/ml in hIL-10/hLF, whereas 1.4 mg/ml in hIL-10/hTPO. Ihe downregulation of hTPO to half level of singles' was observed in double transgenic mice. The possible reason why hTPO co-expressed might lead to down-regulation of another human protein was discussed. These results suggested that double transgenic generated by crossbreeding between two singles' could be useful system for bioreactor.

Introduction of tmie Gene Can Recover the Hearing Impairment and Abnormal Behavior in the Circling Mouse

  • Shin, Mi-Jung;Park, Seo-Jin;Pare, Hum Dai;Ryoo, Zae-Young
    • Reproductive and Developmental Biology
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    • v.34 no.2
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    • pp.63-71
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    • 2010
  • The spontaneous mutant circling mouse (cir/cir) shows a circling behavior and hearing loss. We produced transgenic mice overexpressing transmembrane inner ear (tmie) gene, the causative gene, for the phenotypic rescue of the circling mouse. Through the continuous breeding with circling mice, the cir/cir homozygous mice carrying the transgene (cir/cir-tg) were produced. The rescued cir/cir-tg mice were able to swim in the water with proper orientation and did not show any circling behavior like wild type mice. Western blot and immunohistochemical analysis exhibited that the transgenic tmie was expressed in the inner ear. Inner and outer hair cells were recovered in the cochlea and spiral ganglion neurons were also recovered in the rescued mice. Auditory brainstem response (ABR) test demonstrated that the cir/cir-tg mice are able to respond to sound. This study demonstrates that tmie transgene can recover the hearing impairment and abnormal behavior in the circling mouse.

Fidelity of Transgene Transmission and Expression in the Transgenic Mice

  • Zheng, Z. Y.;Y. M. Han;Y. K. Kang;K. B. Oh;W. J. Shin;Lee, K. K.
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.89-89
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    • 2002
  • In this study, we examined transmission efficiency and expression level of the transgenes in the transgenic mice. The transgenic lines secreting a considerable amount of human lactoferrin(LF) thrombopoietin(TPO), interleukin-10(IL-10) into their milk were subjected to access the inheritance and maintenance of transgenic phenotype. They were bred through three generations. The transmission frequency for each generations(F9, F10, F11) of 3 lines was 38.03±10.43%(13/35), 48.33±3.76%(19/39) and 31.83±8.88%(9/28) in the LF line, 51.33±18.98%(20/38), 63.70±35.71%(12/20) and 29.57± 15.05%(8/26) in the TPO line, 38.27±17.74%(15/37), 47.47±29.88%(14/28) and 50.87±5.85%(14/28) in the IL-10 line, respectively. (omitted)

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Establishment of Transgenic Mouse with the E6 and E7 Genes of Human Papillomavirus Type 16 (인간 Papillomavirus의 E6, E7 유전자를 이용한 Transgenic Mouse의 확립)

  • Hwang, Yong-Il;Lee, Seung-Cheol;Kim, Hyun-Su
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.115-120
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    • 1996
  • Human papillomavirus (HPV), especially type 16 and 18, has been closely associated with carcinomas and uterine cevical cancer, recently. From in vitro assays, E6 and E7 genes of HPV16 are closely linked with transformation of cell lines of rodent fibroplasts. However, the transforming activity of E6 and E7 genes of HPV type 16 in vivo has not been fully elucidated. For explaining this mechanism, we prepared a expression system with the promoter of mouse mammary tumorvirus long terminal repeat and E6E7's open reading frames. This expression system was introduced in rodent cell lines, No. 7, 3Y1 and shown normal transforming abilities. And, we produced transgenic mice with E6, E7 expression system. These transgenic mice were confirmed from Southern blot analysis. One male of them was observed enlargement of the testis after 5 months postdelivery.

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Effects of Garlic on Uncoupling Protein 2 (UCP2) Transcriptional Regulation in Metabolic Tissues of UCP2 Transgenic Mice Fed on a High-Fat Diet (마늘이 고지방 식이를 섭취한 UCP2 형질전환 마우스의 대사성 조직에서 UCP2 전사 조절에 미치는 영향)

  • Lee, Mak-Soon;Lee, Seohyun;Shin, Yoonjin;Jung, Sunyoon;Park, Seonyoung;Kim, Yangha
    • The Korean Journal of Food And Nutrition
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    • v.30 no.3
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    • pp.531-538
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    • 2017
  • This study was performed to investigate the effects of garlic on uncoupling protein 2 (UCP2) transcriptional regulation of UCP2-luciferase transgenic mice fed on a high fat diet to induce obesity. To examine the transcriptional regulation of UCP2, we generated transgenic mice with a UCP2 promoter (-1,830/+30 bp) containing luciferase as a reporter gene. UCP2-luciferase transgenic mice were fed a 45% high-fat diet for 8 weeks to induce obesity. Subsequently, mice were maintained on either a high-fat control diet (TG-CON), or high-fat diets supplemented with 2% (TG-GL2) or 5% (TG-GL5) garlic for a further 8 weeks. Dietary garlic reduced body weight and energy efficiency ratio in the TG-GL5 group, compared to the TG-CON group. Furthermore, garlic supplementation significantly decreased white adipose tissue fat mass and plasma levels of triglycerides, total cholesterol, and leptin in the TG-GL2 and TG-GL5 groups, compared to the TG-CON group. Specifically, UCP2 promoter activity in metabolic tissues such as liver, white adipose tissue, brown adipose tissue, and skeletal muscle was increased by garlic supplementation. These results suggest that dietary garlic was partially associated with an increase of UCP2 transcriptional activity in metabolic tissues for decreasing obesity.