• 제목/요약/키워드: Gene Targeting

검색결과 483건 처리시간 0.029초

한국재래흑염소 수정란의 이식;형질전환 흑염소 생산을 위한 수정란의 채취와 이식 (Embryo transfer in Korean Native Black Goat;Embryo recovery and transfer for the production of transgenic goat)

  • 신상태
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2000년도 춘계심포지움
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    • pp.64-75
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    • 2000
  • During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period (1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor (hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

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MicroRNA-328 Inhibits Proliferation of Human Melanoma Cells by Targeting TGFB2

  • Li, Jing-Rong;Wang, Jian-Qin;Gong, Qing;Fang, Rui-Hua;Guo, Yun-Long
    • Asian Pacific Journal of Cancer Prevention
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    • 제16권4호
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    • pp.1575-1579
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    • 2015
  • Some microRNAs (miRNAs) have been shown to act as oncogenes or tumor suppressor genes in human melanomas. miR-328 is upregulated in blood cells of melanoma patients compared to in healthy controls. This suggests a role for miR-328 in melanoma that warrants investigation. In this study, we demonstrated miR-328 levels to be dramatically decreased in human melanoma cell lines. Moreover, forced expression of miR-328 inhibited proliferation and induced G1-phase arrest of the SK-MEL-1 melanoma cell line. We identified TGFB2 as a direct target gene for miR-328 using a fluorescent reporter assay and western blotting. Levels of TGFB2 were dramatically increased in human melanoma cell lines and were inversely correlated with the miR-328 expression level. Our findings provide new insights into the mechanisms of human melanoma development, indicating that miR-328 has therapeutic potential for this disease.

MicroRNA-100 Resensitizes Resistant Chondrosarcoma Cells to Cisplatin through Direct Targeting of mTOR

  • Zhu, Zhe;Wang, Cun-Ping;Zhang, Yin-Feng;Nie, Lin
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권2호
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    • pp.917-923
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    • 2014
  • Chondrosarcomas are malignant cartilage-forming tumors of bone which exhibit resistance to both chemotherapy and radiation treatment. miRNAs have been well demonstrated to regulate gene expression and play essential roles in a variety of biological processes, including proliferation, differentiation, migration, cell cycling and apoptosis. In this study, we obtained evidence that miR-100 acts as a tumor suppressor in human chondrosarcomas. Interestingly, cisplatin resistant chondrosarcoma cells exhibit decreased expression of miR-100 compared with parental cells. In addition, we identified mTOR as a direct target of miR-100. Overexpression of miR-100 complementary pairs to the 3' untranslated region (UTR) of mTOR, resulted in sensitization of cisplatin resistant cells to cisplatin. Moreover, recovery of the mTOR pathway by overexpression of S6K desensitized the chondrosarcoma cells to cisplatin, suggesting the miR-100-mediated sensitization to cisplatin dependent on inhibition of mTOR. In summary, the present studies highlight miR-100 as a tumor suppressor in chondrosarcoma contributing to anti-chemoresistance. Overexpression of miR-100 might be exploited as a therapeutic strategy along with cisplatin-based combined chemotherapy for the treatment of clinical chondrosarcoma patients.

Insights into the Diverse Roles of miR-205 in Human Cancers

  • Orang, Ayla Valinezhad;Safaralizadeh, Reza;Feizi, Mohammad Ali Hosseinpour
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권2호
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    • pp.577-583
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    • 2014
  • The recent discovery of tiny microRNAs (miRNAs) has brought about awareness of a new class of regulators of diverse pathways in many physiological and pathological processes, such as tumorigenesis. They modulate gene expression by targeting plethora of mRNAs, mostly reducing the protein yield of a targeted mRNA. With accumulation of information on characteristics of miR-205, complex and in some cases converse roles of miR-205 in tumor initiation, progression and metastasis are emerging. miR-205 acts either as an oncogene via facilitating tumor initiation and proliferation, or in some cases as a tumor suppressor through inhibiting proliferation and invasion. The aim of this review is to discuss miR-205 roles in different types of cancers. Given the critical effects of deregulated miR-205 on processes involved in tumorigenesis, they hold potential as novel therapeutic targets and biomarkers.

A new function of glucocorticoid receptor: regulation of mRNA stability

  • Park, Ok Hyun;Do, Eunjin;Kim, Yoon Ki
    • BMB Reports
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    • 제48권7호
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    • pp.367-368
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    • 2015
  • It has long been thought that glucocorticoid receptor (GR) functions as a DNA-binding transcription factor in response to its ligand (a glucocorticoid) and thus regulates various cellular and physiological processes. It is also known that GR can bind not only to DNA but also to mRNA; this observation points to the possible role of GR in mRNA metabolism. Recent data revealed a molecular mechanism by which binding of GR to target mRNA elicits rapid mRNA degradation. GR binds to specific RNA sequences regardless of the presence of a ligand. In the presence of a ligand, however, the mRNA-associated GR can recruit PNRC2 and UPF1, both of which are specific factors involved in nonsense-mediated mRNA decay (NMD). PNRC2 then recruits the decapping complex, consequently promoting mRNA degradation. This mode of mRNA decay is termed "GR-mediated mRNA decay" (GMD). Further research demonstrated that GMD plays a critical role in chemotaxis of immune cells by targeting CCL2 mRNA. All these observations provide molecular insights into a previously unappreciated function of GR in posttranscriptional regulation of gene expression. [BMB Reports 2015; 48(7): 367-368]

Modulation of the Metal(loid) Specificity of Whole-Cell Bioreporters by Genetic Engineering of ZntR Metal-Binding Loops

  • Kim, Hyojin;Jang, Geupil;Kim, Bong-Gyu;Yoon, Youngdae
    • Journal of Microbiology and Biotechnology
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    • 제30권5호
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    • pp.681-688
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    • 2020
  • Bacterial cell-based biosensors, or whole-cell bioreporters (WCBs), are an alternative tool for the quantification of hazardous materials. Most WCBs share similar working mechanisms. In brief, the recognition of a target by sensing domains induces a biological event, such as changes in protein conformation or gene expression, providing a basis for quantification. WCBs targeting heavy metal(loid)s employ metalloregulators as sensing domains and control the expression of genes in the presence of target metal(loid) ions, but the diversity of targets, specificity, and sensitivity of these WCBs are limited. In this study, we genetically engineered the metal-binding loop (MBL) of ZntR, which controls the znt-operon in Escherichia coli. In the MBL of ZntR, three Cys sites interact with metal ions. Based on the crystal structure of ZntR, MBL sequences were modified by site-directed mutagenesis. As a result, the metal-sensing properties of WCBs differed depending on amino acid sequences and the new selectivity to Cr or Pb was observed. Although there is room for improvement, our results support the use of currently available WCBs as a platform to generate new WCBs to target other environmental pollutants including metal(loid)s.

Silibinin Inhibits LPS-Induced Macrophage Activation by Blocking p38 MAPK in RAW 264.7 Cells

  • Youn, Cha Kyung;Park, Seon Joo;Lee, Min Young;Cha, Man Jin;Kim, Ok Hyeun;You, Ho Jin;Chang, In Youp;Yoon, Sang Pil;Jeon, Young Jin
    • Biomolecules & Therapeutics
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    • 제21권4호
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    • pp.258-263
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    • 2013
  • We demonstrate herein that silibinin, a polyphenolic flavonoid compound isolated from milk thistle (Silybum marianum), inhibits LPS-induced activation of macrophages and production of nitric oxide (NO) in RAW 264.7 cells. Western blot analysis showed silibinin inhibits iNOS gene expression. RT-PCR showed that silibinin inhibits iNOS, TNF-${\alpha}$, and $IL1{\beta}$. We also showed that silibinin strongly inhibits p38 MAPK phosphorylation, whereas the ERK1/2 and JNK pathways are not inhibited. The p38 MAPK inhibitor abrogated the LPS-induced nitrite production, whereas the MEK-1 inhibitor did not affect the nitrite production. A molecular modeling study proposed a binding pose for silibinin targeting the ATP binding site of p38 MAPK (1OUK). Collectively, this series of experiments indicates that silibinin inhibits macrophage activation by blocking p38 MAPK signaling.

An Arabidopsis Homologue of Human Seven-in-Absentia-interacting Protein Is Involved in Pathogen Resistance

  • Kim, Youn-Sung;Ham, Byung-Kook;Paek, Kyung-Hee;Park, Chung-Mo;Chua, Nam-Hai
    • Molecules and Cells
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    • 제21권3호
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    • pp.389-394
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    • 2006
  • Human seven-in-absentia (SIAH)-interacting protein (SIP) is a component of the E3 ligase complex targeting beta-catenin for destruction. Arabidopsis has one SIP protein (AtSIP) with 32% amino acid sequence identity to SIP. To investigate the functions of AtSIP, we isolated an atsip knockout mutant, and generated transgenic plants overexpressing AtSIP. The growth rates and morphologies of the atsip and transgenic plants were indistinguishable from those of wild type. However, atsip plants were more susceptible to Pseudomonas syringae infection, and the transgenic plants overexpressing AtSIP were more resistant. Consistent with this, RNA blot analysis showed that the AtSIP gene is strongly induced by wounding and hydrogen peroxide treatment. In addition, when plants were infected with P. syringae, AtSIP was transiently induced prior to PR-1 induction. These observations show that Arabidopsis AtSIP plays a role in resistance to pathogenic infection.

한국재래흑염소 수정란의 이식: 형질전환 흑염소 생산을 위한 수정란의 채취와 이식 (Embryo transfer in Korean Native Black Goat: Embryo recovery and transfer for the production of transgenic goat)

  • 신상태
    • 한국수정란이식학회:학술대회논문집
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    • 한국수정란이식학회 2000년도 수정란의 위생적 처리·검사 및 특수가축의 수정란이식 기술 심포지움
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    • pp.64-75
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    • 2000
  • During the last three decades considerable advances has been made in goat embryo production and transfer technology. The Korean native black goat is the most useful domestic ruminant in this country for biological investigation and application because it has a lot of merits such as relatively short generation period(1 vs 2 year for a cow), easy of handling, well adaptation, high fertility, convenient and inexpensive. This article covers the methods of superovulation, estrus synchronization, embryo collection and transfer techniques, pregnancy diagnosis and subsequent pregnancy and kidding rates for the production of transgenic Korean native black goats. More than one hundred goat kids have been produced as a result of our transgenic goat project via microinjection of foreign gene into pronuclei, in vitro culture, transfer of various stages of fresh and frozen-thawed microinjected embryos into oviducts or uteri of recipient does. We have got two transgenic goats carrying a transgene targeting the expression of recombinant human granulocyte colony stimulating factor(hG-CSF) to the mammary gland so far. Since collection and transfer of embryos in this species is usually accomplished by laparotomy, exteriorization of the reproductive tract for surgical embryo collection leads to the formation of post-operative adhesions. Nonsurgical or laparoscopic technique to reduce adhesions from repeated surgeries has great advantages in improving embryo production and transfer especially from valuable donors. We will discuss this later.

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Galaxy overdensities at intermediate to high redshift

  • 강유진;임명신
    • 천문학회보
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    • 제36권2호
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    • pp.52.2-52.2
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    • 2011
  • We searched and studied galaxy overdensities at 0.6 < z < 4.5 in the areas of two GOODS fields. These overdensities are identified by running top-hat filters on the two dimensional spatial distribution of two galaxy samples - a K-band limited, photometric redshift sample at 0.6 < z < 4.5, and BVz-color selected sample targeting overdensities at z ~ 3.7 and z ~ 4.0. Here photometric redshifts are derived from multi-wavelength data ranging from U-band through 8 micron band of the Spitzer. We find 52 overdensities with significances of 3.5-7 ${\sigma}$. The derived masses found to be a > $10^{13}\;M_{\odot}$ with the comoving number density of a few ${\times}10^{-6}\;Mpc^{-3}$ at z ~ 3. In order to understand the high number density of massive overdensities at high redshift, we carried out an analysis of galaxy overdensities using the mock galaxy catalog based on Millennium simulation selected in the same way as the analysis of the observational data. In the simulation, we find 650 galaxy overdensities with a 3.5 ${\sigma}$ detection threshold over $2^{{\circ}2}$ sky field. The number density of the very massive overdensities (M > $10^{14}\;M_{\odot}$) in simulation shows a similar trend with the observation. We further discuss implications of our results.

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