• Title/Summary/Keyword: oligonucleotides

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MicroRNA-802 Promotes Osteosarcoma Cell Proliferation by Targeting p27

  • Cao, Zhong-Qing;Shen, Zan;Huang, Wei-Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.12
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    • pp.7081-7084
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    • 2013
  • MicroRNAs have been demonstrated to regulate proliferation and apoptosis in many types of cancers, but biological functions in osteosarcomas remain relatively unknown. Here, we found expression of miR-802 to be up-regulated in osteosarcoma tissues in comparison with adjacent normal tissues. Enforced expression of miR-802 was able to promote cell proliferation in U2OS and MG63 cells, while miR-802 antisense oligonucleotides (antisense miR-802) inhibited cell proliferation. At the molecular level, our results further revealed that expression of p27, a negative cell-cycle regulator, was negatively regulated by miR-802. Therefore, the data reported here indicate that miR-802 is an important regulator in osteosarcoma, our findings contributing to a better understanding of important mis-regulated miRNAs in this tumour type.

Development of DNA Chip Microarrayer

  • Yoon, Sung-Ho;Choi, Jong-Gil;Lee, Sang-Yup
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.21-26
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    • 2000
  • A microarrayer system was developed mainly for manufacturing DNA chips. The 3-axis robot was designed to automatically collect samples from 96-or 384-well microtiter plates using up to 16 simultaneously moving pens and to deposit them on a surface-modified slide glass. This is followed by a wash/dry operation in a clean station. The cycle is repeated with a new set of samples, This system can deposit cDNA or oligonucleotides with spot intervals of $150{\;}\mu\textrm{m}$ and the spot size of $80\mu\textrm{m}$, thus allowing a high density DNA chip containing about 5,000 spots per $\textrm{cm}^2$. The entire procedure is controlled by the Visual C++ program that was written in our laboratory by using a personal computer with Pentium 100 CPU.

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Genetic Differences within and between Populations of Korean Catfish (S. asotus) and Bullhead (P. fulvidraco) Analysed by RAPD-PCR

  • Yoon, Jong-Man;Kim, Jong-Yeon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2004.05a
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    • pp.321-322
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    • 2004
  • Of the 20 arbitrarily chosen primers, six oligonucleotides decamer primers were used on the basis of the number of the polymorphisms generated in catfish (Silurus asotus) from Yesan and bullhead (Pseudobagrus fulvidraco) from Dangjin in Korea. Six primers were used generating a total of 602 scorable bands in catfish and 195 in bullhead population, respectively, ranging in size of DNA fragments from less than approximately 100 to larger than 2,000 base pairs (bp). Six primers yielded 199 polymorphic fragments (33.1 %) in catfish and 47 (24 %) in bullhead, respectively. In the present study, a total of 328 common fragments (an average of 54.7 per prime.) were observed in catfish population, whereas 84 (an average of 14.0 per prime.) in bullhead. The total number of specific fragments in catfish and bullhead population were 76and 64, respectively.

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MicroRNA-16 Inhibits Bladder Cancer Proliferation by Targeting Cyclin D1

  • Jiang, Qi-Quan;Liu, Bin;Yuan, Tao
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.7
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    • pp.4127-4130
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    • 2013
  • MicroRNA-16 (miR-16) has been demonstrated to regulate proliferation and apoptosis in many types of cancers, but its biological function in bladder cancer remains unknown. Here, we found expression of miR-16 to be downregulated in bladder cancer in comparison with the adjacent normal tissues. Enforced expression of miR-16 was able to inhibit cell proliferation in TCHu-1 cells, in line with results for miR-16 antisense oligonucleotides (antisense miR-16). At the molecular level, our results further revealed that cyclin D1 expression was negatively regulated by miR-16. Therefore, the data reported here demonstrate that miR-16 is an important regulator in bladder cancer, which will contribute to better understanding of important mis-regulated miRNAs.

HIV Integrase Inhibitory Activity of Agastache regosa

  • Kim, Hye-Kyung;Lee, Hyeong-Kyu;Shin, Cha-Gyun;Huh, Hoon
    • Archives of Pharmacal Research
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    • v.22 no.5
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    • pp.520-523
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    • 1999
  • We have been screening anti-HIV integrase compounds from Korean medicinal plants by using an in vitro assay system which is mainly composed of recombinant human immunodeficency virus type 1 integrase and radiolabeled oligonucleotides. From the above screening, the aqueous methanolic extract of the roots of Agastache rugosa exhibited a significant activity. Bioactivity-guided chromatographic fractionation of the methanolic extract resulted in the isolation of rosmarinic acid. The structure of the compound was determined by spectroscopic data and by the comparison with the reported values. The $IC_{50}$ of the rosmarinic acid was approximately $10{\mu}g/ml$ against HIV integrase.

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Glial Cell-specific Regulation of the JC virus Early Promoter by Silencer and DNA Methylation (Silencer 및 DNA methylation에 의한 JC virus early promoter의 뇌교세포 특이적인 조절)

  • 김희선;우문숙
    • YAKHAK HOEJI
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    • v.46 no.2
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    • pp.143-148
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    • 2002
  • The human polyomavirus JC virus is the etiologic agent of progressive multifocal leukoencephalopathy (PML). The JC virus early promoter directs cell-specific expression of the viral replication factor large T antigen, thus transcriptional regulation constitutes a major mechanism of glial tropism in PML. Here we found that pentanucleotide sequence immediately upstream of the TATA sequence functions as a cell-specific silencer in the JC virus transcription. In vitro binding studies showed that synthetic oligonucleotides spanning a pentanucleotide sequence, designated "oligo 2", interacts with nuclear proteins from non-glial cells in a cell-specific manner. Furthermore, the sequence preferentially repressed the heterologous thymidine kinase promoter activity in non-glial cells. We also tested whether JC virus transcription is controlled by DNA methylation. Transient transfection of in vitro methylated JC virus promoter abolished transcription in both the glial and non-glial cells. The repression fold was much larger in glial cells than in non-glial cells. Taken together, this finding suggests that glial cell-specific expression of the JC virus is controlled by DNA methylation as well as cell-specific silencers.

Functional Analysis of RAD4 Gene Required for Nucleotide Excision Repair of UV-induced DNA Damage in Saccharomyces cerevisiae

  • Park, Sang Dai;Park, In Soon
    • Animal cells and systems
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    • v.6 no.4
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    • pp.311-315
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    • 2002
  • The RAD4 gene is essential for nucleotide excision repair in Saccharomyces cerevisiae. It has been known that the deduced amino acid sequence of Rad4 protein contains three DNA-dependent ATPase/helicase motifs. To determine the biochemical activities and functional role of RAD4 the Rad4 protein was expressed and purified. Immunoblot analysis showed a specific band of 21 kDa, which was well-matched with the size of open reading frame of the RAD4 gene. The purified Rad4 protein had no detectable helicase activity. However, the protein could interact with double stranded oligonucleotides, as judged by mobility shift assay. This result suggests that the Rad4 protein is a DNA binding protein.

Nanoparticle-based Detection Technology for DNA Analysis

  • Park, Hyun-Gyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.221-226
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    • 2003
  • With the current rapid development of nanotechnology and synthesis technology for designed oligonucleotides or oligonucleotide-modified nanoparticle conjugates, the combined strategies have become one of the most valuable methods in detection technology for DNA analysis. Using the uniquely recognizable interactions of pre-designed DNA molecules in assembling nanoparticles, various novel approaches have been recently developed towards detecting specific DNA sequences. Here we describe the key fundamentals and issues of this promising strategies ranging from the initial findings of rationally designed DNA-based assembly of nanoparticles to the extended chip-based detection system. Some limitations of these new strategies and possible approaches will be also discussed for the practical application in the area of DNA microarray detection.

Points To Consider in Development and Evaluation of the Gene Therapy Products in Korea (국내 유전자치료 의약품 개발 및 평가 시 고려해야 할 사항)

  • Kim, Jin-Seok
    • Journal of Pharmaceutical Investigation
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    • v.30 no.2
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    • pp.139-143
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    • 2000
  • Gene therapy is becoming a very promising and feasible medical intervention as the understanding of human diseases extends to their molecular levels. Since the first US Food and Drug Administration (FDA)-approved human gene therapy protocol was approved in 1990, over 300 human clinical trial protocols had been approved worldwide so far. Even though some of the domestic gene therapy clinical trials also proved promising and more are awaiting, it should be emphasized that many safety aspects as well as effectiveness aspects should be considered during the development process. Moreover, there seems to be less restricted guidelines from the National Control Authority (NCA) in initiating human clinical trials. This article is intended to suggest some basis and points to consider in the development and evaluation of gene therapy products including antisense oligonucleotides pharmaceuticals.

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NMR Study on Thermal Stability of the Double Helical Structures of d(CGAATTCG)2, d(CGTATACG)2 and their berenil complexes

  • Kim, Eun-Hee;Hong, Seok-Joo;Huh, Sung-Ho
    • Journal of the Korean Magnetic Resonance Society
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    • v.8 no.2
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    • pp.96-107
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    • 2004
  • We prepared two oligonucleotides containing same base pairing, but different base sequence in the middle region, d(CGAATTCG) and d(CGTATACG). NMR and UV absorbance data represented that such variation in base sequence could cause a significant difference in melting temperature and dynamics between d(CGAATTCG)$_2$ and d(CGTATACG)$_2$ duplexes, which are regarded to be associated with the stacked structure and the width of the minor groove of them. The latter showed poor stability compared to the former, because of poor stacking of bases. And berenil could bind to the minor groove of d(CGAATTCG)$_2$ which is relatively narrow, more strongly than d(CGTATACG)$_2$ and this gave rise to large improvement in thermal stability of the d(CGAATTCG)$_2$ duplex, compared to d(CGTATACG)$_2$.

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