• Title/Summary/Keyword: reporter system

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A New Reporter Vector System Based on Flow-Cytometry to Detect Promoter Activity

  • Jung, Sun-Do;Choi, Ji-Hye;Hong, Chang-Wan;Lee, Hyun-Ji;Park, Yoon-Kyung;Shin, Jung-Hoon;Park, Jae-Won;Park, Se-Ho
    • IMMUNE NETWORK
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    • v.9 no.6
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    • pp.243-247
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    • 2009
  • In this study, we report the development of a new dual reporter vector system for the analysis of promoter activity. This system employs green fluorescence emitting protein, EGFP, as a reporter, and uses red fluorescence emitting protein, DsRed, as a transfection control in a single vector. The expression of those two proteins can be readily detected via flow cytometry in a single analysis, with no need for any further manipulation after transfection. As this system allows for the simultaneous detection of both the control and reporter proteins in the same cells, only transfected cells which express the control protein, DsRed, can be subjected to promoter activity analysis, via the gating out of all un-transfected cells. This results in a dramatic increase in the promoter activity detection sensitivity. This novel reporter vector system should prove to be a simple and efficient method for the analysis of promoter activity.

Study of Synergy Effects of Collaboration in News Program between TV Reporter & Producer - Focusing on KBS 2TV - (방송뉴스제작에 있어서 기자.PD 협업의 시너지효과 탐구 - KBS 2TV <뉴스투데이>를 중심으로 -)

  • Hong, Kyung-Soo
    • The Journal of the Korea Contents Association
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    • v.11 no.4
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    • pp.164-176
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    • 2011
  • To get an implication for desirable direction for Collaboration between Reporter & Producer, I found that had 4 modes of collaboration such as, Producer System, Reporter System, Double Reporting System, Co-Work System, and each was distinguished by field cover, on-mic, editing, writing. Producer system and Reporter system got no chemical synergy because of simple array of single reports due to physical assemble. The achievements of collaboration was birth of new news which had different duration, visual, sound & perspective. It also offered fertile soil for new program and genre literacy of 8 o'clock news. The success factor of collaboration was surprisingly turned out to be personnel rather than system. The balance of reporter & producer was also good factor. But the wall between 2 groups was so high that, after disappearance of extraordinary leader, system began to crumble and at last program was repealed.

Porcine OCT4 reporter system as a tool for monitoring pluripotency states

  • Kim, Seung-Hun;Lee, Chang-Kyu
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.4
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    • pp.175-182
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    • 2021
  • Pluripotent stem cells could self-renew and differentiate into various cells. In particular, porcine pluripotent stem cells are useful for preclinical therapy, transgenic animals, and agricultural usage. These stem cells have naïve and primed pluripotent states. Naïve pluripotent stem cells represented by mouse embryonic stem cells form chimeras after blastocyst injection. Primed pluripotent stem cells represented by mouse epiblast stem cells and human embryonic stem cells. They could not produce chimeras after blastocyst injection. Populations of embryonic stem cells are not homogenous; therefore, reporter systems are used to clarify the status of stem cells and to isolate the cells. For this reason, studies of the OCT4 reporter system have been conducted for decades. This review will discuss the naïve and primed pluripotent states and recent progress in the development of porcine OCT4 reporter systems.

Moving Object Tracing System using Network-connected CCTV and Smartphone (네트워크 CCTV와 스마트 단말기를 연동한 이동체 추적 시스템)

  • Lim, Seungkyun;Lee, Sangwook
    • The Journal of the Korea Contents Association
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    • v.13 no.12
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    • pp.519-529
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    • 2013
  • This paper introduces a moving object tracing system using network-connected CCTV and smartphone. In an emergency situation, a smartphone of the reporter gains its GPS information and sends that to the central server. The central server stores received GPS information as a dangerous area in the database and keeps sending the order of tracing the reporter to a network-connected CCTV which can film the reporter. At the same time, the central server sends pictures of the reporter to his or her family and related organization in order to handle the emergency situation as soon as possible. In addition, when a reporter want to know the risk around destination, the central server informs dangerous areas to the reporter by using smart phone application and database of a danger spot.

Production of Mutated Porcine Embryos Using Zinc Finger Nucleases and a Reporter-based Cell Enrichment System

  • Koo, Ok Jae;Park, Sol Ji;Lee, Choongil;Kang, Jung Taek;Kim, Sujin;Moon, Joon Ho;Choi, Ji Yei;Kim, Hyojin;Jang, Goo;Kim, Jin-Soo;Kim, Seokjoong;Lee, Byeong-Chun
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.3
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    • pp.324-329
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    • 2014
  • To facilitate the construction of genetically-modified pigs, we produced cloned embryos derived from porcine fibroblasts transfected with a pair of engineered zinc finger nuclease (ZFN) plasmids to create targeted mutations and enriched using a reporter plasmid system. The reporter expresses RFP and eGFP simultaneously when ZFN-mediated site-specific mutations occur. Thus, double positive cells ($RFP^+/eGFP^+$) were selected and used for somatic cell nuclear transfer. Two types of reporter based enrichment systems were used in this study; the cloned embryos derived from cells enriched using a magnetic sorting-based system showed better developmental competence than did those derived from cells enriched by flow cytometry. Mutated sequences, such as insertions, deletions, or substitutions, together with the wild-type sequence, were found in the cloned porcine blastocysts. Therefore, genetic mutations can be achieved in cloned porcine embryos reconstructed with ZFN-treated cells that were enriched by a reporter-based system.

Imaging Gene Expression (유전자 발현 영상기법)

  • Lee, Kyung-Han
    • The Korean Journal of Nuclear Medicine
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    • v.34 no.1
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    • pp.1-9
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    • 2000
  • The rapid progress of molecular genetic methods over the past two decades has necessitated the development of methods to detect and quantify genetic activity within living bodies. Reporter genes provide a rapid and convenient tool to monitor gene expression by yielding a readily measurable phenotype upon expression when introduced into a biological system. Conventional reporter systems, however, are limited in their usefulness for in vivo experiments or human gene therapy because of its invasive nature which requires cell damage before assays can be performed. This offers an unique opportunity for nuclear imaging techniques to develope a novel method for imaging both the location and amount of gene expression noninvasively. Current developments to achieve this goal rely on utilizing either reporter enzymes that accumulate radiolabeled substrates or reporter receptors that bind specific radioligands. This overview includes a brief introduction to the background for such research, a summary of published results, and an outlook for future directions.

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Cytokine Reporter Mouse System for Screening Novel IL12/23 p40-inducing Compounds

  • Im, Wooseok;Kim, Hyojeong;Yun, Daesun;Seo, Sung-Yum;Park, Se-Ho;Locksley, Richard M.;Hong, Seokmann
    • Molecules and Cells
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    • v.20 no.2
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    • pp.288-296
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    • 2005
  • Cytokines interleukin (IL) 12 and 23 play critical roles in linking innate and adaptive immune responses. They are members of heterodimeric cytokines, sharing a subunit p40. Although IL12/23 p40 is mainly induced in macrophages and dendritic cells (DCs) after stimulation with microbial Toll-like receptor ligands, methods to monitor the cells that produce IL12/23 p40 in vivo are limited. Recently, the mouse model to track p40-expressing cells with fluorescent reporter, yellow fluorescent protein, has been developed. Macrophages and DCs from these mice faithfully reported p40 induction using the fluorescent marker. Here we took advantage of these reporter mice to screen bio-compounds for p40-inducing activity. After screening hundreds of compounds, we found several extracts inducing IL12/23 p40 gene expression. Treatment of DCs with these extracts induced the expression of MHC class II and co-stimulatory molecules, which implies that these might be useful as adjuvants. Next, the in vivo target immune cells of candidate compounds were examined. The reporter system can be useful to identify cells producing IL12 or IL23 in vivo as well as in vitro. Thus, our cytokine reporter system proved to be a valuable reagent for screening for immunostimulatory molecules and identification of target cells in vivo.

Establishment of Reporter Cell Lines that Monitor Activities of Hypoxia Inducible Factor-1, P53 and Nur77 for Assessment of Carcinogenicity (저산소유도인자 HIF-1, 암 억제인자 P53과 고아 핵수용체 Nur77의 발현을 지표로 하는 발암독성예측 세포주의 개발)

  • Hong, Il;Seo, Hee-Won;Lee, Min-Ho;Kim, Ji-Won;Chung, Jin-Ho;Lee, Byung-Hoon;Lee, Mi-Ock
    • Toxicological Research
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    • v.23 no.3
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    • pp.231-238
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    • 2007
  • Evaluation of potentials of chemicals to alter expression of genes that are involved in carcinogenesis may serve useful tools in toxicological research. In this investigation, we developed reporter cell lines that expressed luciferase in response to transactivation of hypoxia inducible factor-1, P53 tumor suppressor and Nur77 of which roles have been well established in cancer development and progression. Whereas these reporter cell lines displayed low constitutive backgrounds, the reporter activities were significantly enhanced in response to $desferriosamine/CoCl_2$, adriamycin or 6-mercaptopurine, which are hypoxia mimicking chemicals, P53 activator or Nur77 inducer, respectively. The activation of the reporter was time- and dose-dependent. Known tumor initiators and promoters, such as phorbol 12-myristate 13-acetate and phorbol 12, 13-dicaprinate induced the reporter activity at as low as 10nM in these stable cell lines. Further, known anti-tumor promoters, such as ascorbic acid and ${\beta}-carotene$ repressed the reporter activities. These results indicate that our stable reporter cell lines could serve as a useful system for rapid assessment of carcinogenicity of toxic chemicals.

The Rat Myosin Light Chain Promoter-Driven DsRed Reporter System Allows Specific Monitoring of Bone Marrow Mesenchymal Stem Cell- Derived Cardiomyocytes

  • Choi, Seung-Cheol;Lim, Do-Sun
    • Reproductive and Developmental Biology
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    • v.32 no.1
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    • pp.21-25
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    • 2008
  • Bone marrow mesenchymal stem cells (BMMSCs) have the capacity for self-renewal and differentiation into a variety of cell types. They represent an attractive source of cells for gene and cell therapy. The purpose of this study is to direct the specific expression of the DsRed reporter gene in $Sca-1^+$ BMMSCs differentiated into a cardiomyogenic lineage. We constructed the prMLC-2v-DsRed vector expressing DsRed under the control of the 309 tp fragment of the rat MLC-2v 5'-flanking region. The specific expression of the DsRed reporter gene under the transcriptional control of the 309 bp fragment of the rat MLC-2v promoter was tested in 5-azacytidine healed-$Sca-1^+$ BMMSCs over 2 weeks after the prMLC-2v-DsRed transfection. The prMLC-2v-DsRed was specifically expressed in the $Sca-1^+$ BMMSCs with cardiomyogenic lineage differentiation and it demonstrates that the 309 bp sequences of the rat MLC-2v 5'-flanking region is sufficient to confer cardiac specific expression on a DsRed reporter gene. The cardiac-specific promoter-driven reporter vector provides an important tool for the study of stem cell differentiation and cell replacement therapy in ischemic cardiomyopathy.

Development of a Reporter System for In Vivo Monitoring of γ-Secretase Activity in Drosophila

  • Hong, Young Gi;Roh, Seyun;Paik, Donggi;Jeong, Sangyun
    • Molecules and Cells
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    • v.40 no.1
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    • pp.73-81
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    • 2017
  • The ${\gamma}$-secretase complex represents an evolutionarily conserved family of transmembrane aspartyl proteases that cleave numerous type-I membrane proteins, including the ${\beta}$-amyloid precursor protein (APP) and the receptor Notch. All known rare mutations in APP and the ${\gamma}$-secretase catalytic component, presenilin, which lead to increased amyloid ${\beta}$-peptide production, are responsible for early-onset familial Alzheimer's disease. ${\beta}$-amyloid protein precursor-like (APPL) is the Drosophila ortholog of human APP. Here, we created Notch- and APPL-based Drosophila reporter systems for in vivo monitoring of ${\gamma}$-secretase activity. Ectopic expression of the Notch- and APPL-based chimeric reporters in wings results in vein truncation phenotypes. Reporter-mediated vein truncation phenotypes are enhanced by the Notch gain-of-function allele and suppressed by RNAi-mediated knockdown of presenilin. Furthermore, we find that apoptosis partly contributes to the vein truncation phenotypes of the APPL-based reporter, but not to the vein truncation phenotypes of the Notch-based reporter. Taken together, these results suggest that both in vivo reporter systems provide a powerful genetic tool to identify genes that modulate ${\gamma}$-secretase activity and/or APPL metabolism.