• Title/Summary/Keyword: Microarrays

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Preparation of Oligonucleotide Arrays with High-Density DNA Deposition and High Hybridization Efficiency

  • Park, Jeong-Won;Jung, Yong-Won;Jung, Young-Hwan;Seo, Jeong-Sun;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.25 no.11
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    • pp.1667-1670
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    • 2004
  • In DNA microarray produced by DNA-deposition technology, DNA-immobilization and -hybridization yields on a solid support are most important factors for its accuracy and sensitivity. We have developed a dendrimeric support using silylated aldehyde slides and polyamidoamine (PAMAM) dendrimers. An oligonucleotide array was prepared through a crosslinking between the dendrimeric support and an oligonucleotide. Both DNAimmobilization and -hybridization yields on the solid support increased by the modification with the dendrimers. The increase of the immobilization and hybridization efficiency seems to result from a threedimensional arrangement of the attached oligonucleotide. Therefore, our dendrimeric support may provide a simple and efficient solution to the preparation of DNA microarrays with high-density DNA-deposition and high hybridization efficiency.

Microarray Analysis of the Gene Expression Profile in Diethylnitrosamine-induced Liver Tumors in Mice

  • Jung Eun-Soo;Park Jung-Duck;Ryu Doug-Young
    • Environmental Mutagens and Carcinogens
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    • v.25 no.4
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    • pp.134-142
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    • 2005
  • Liver cancer is a leading cause of tumor-related mortality, Diethylnitrosamine (DEN) is one of the most extensively studied hepatic carcinogens to date. In this study, the mRNA expression profile in DEN-induced liver tumors in mice was analyzed using DNA microarrays. We report increased expression of genes that participate in hypoxia response, including metallothionein 1 (Mt1), metallothionein 2 (Mt2), fatty acid synthase (Fasn), transferrin (Trf), adipose differentiation-related Protein (AdfP) and ceruloplasmin (CP), as well as those involved in predisposition and development of cancers, such as cytochrome P450 2A5 (Cyp2a5), alpha 2-HS-glycoprotein (Ahsg) and Jun-B oncogene (Junb). The hepatic iron regulatory peptide, hepcidin (Hampl), was downregulated in DEN-stimulated liver tumors. Expression of tumor suppressor genes, such as tripartite motif protein 13 (Trim13), was decreased under these conditions. The data collectively indicate that DEN-induced tumor development can be exploited as a possible model for liver cancer, since this process involves various genes with important functions in hepatic carcinogenesis.

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Prospects for Plant Biotechnology and Bioindustry in the 21s1 Century: Paradigm Shift Driven by Genomics (21세기 식물생명공학과 생물산업의 전망 : 유전체 연구에 의한 Paradigm Shift)

  • Liu, Jang-Ryol;Choi, Dong-Woog;Chung, Hwa-Jee
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.145-150
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    • 2002
  • Biotechnology in the 21st century will be driven by three emerging technologies: genomics, high-throughput biology, and bioinformatics. These technologies are complementary to one another. A large number of economically important crops are currently subjected to whole genome sequencing. Functional genomics for determining the functions of the genes comprising the given plant genome is under progress by using various means including phenotyping data from transgenic mutants, gene expression profiling data from DNA microarrays, and metabolic profiling data from LC/mass analysis. The aim of plant molecular breeding is shifting from introducing agronomic traits such as herbicide and insect resistance to introducing quality traits such as healthful oils and proteins, which will lead to improved and nutritional food and feed products. Plant molecular breeding is also expected to aim to develop crops for producing human therapeutic and industrial proteins.

Changes in Human Gene Expression After Sleep Deprivation

  • Sun, Je Young;Kim, Jong Woo;Yim, Sung-Vin;Oh, Miae;Kang, Won Sub
    • Korean Journal of Biological Psychiatry
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    • v.29 no.1
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    • pp.9-14
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    • 2022
  • Objectives Sleep is fundamental to maintaining homeostatic control and has behavioral and psychological effects on humans. To better understand the function and pathophysiology of sleep, specific gene expressions in reference to sleep deprivation have been studied. In this study, we investigated the gene expression of peripheral blood mononuclear cells after sleep deprivation to better understand the functional consequence of sleep. Methods In eight healthy men, 24 h sleep deprivation was induced. Blood was sampled at 14:00, before and after sleep deprivation. mRNA was isolated and analyzed via microarrays. cDNAs before and after sleep deprivation were coupled to Cy3 or Cy5, respectively, and normalized cDNAs were selected with a ratio greater than two as a significant gene. Results are expressed as mean. Results Among 41174 transcripts, 38852 genes were selected as reliable, and only a small minority (< 1%) of the genes were up-or down-regulated. Total six and eleven genes were selected as significant upregulated and downregulated genes, respectively. Protein tyrosine phosphatase receptor type O was most upregulated (6.9-fold), and low-density lipoprotein receptor-related protein 5-like protein showed the most substantial inhibition (0.06-fold). Conclusions This study showed significant associations between sleep deprivation and the immune system. Acute sleep deprivation affects pathways in proinflammatory cytokines as well as metabolic pathways of glutamate and purine, neurotransmitters related to sleep and wake cycle.

Global trends in regulatory frameworks for animal genome editing in agriculture

  • Dajeong Lim;Inchul Choi
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.4
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    • pp.247-253
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    • 2023
  • Revolutionary advancements, such as the reduction in DNA sequencing costs and genome editing, have transformed biotechnology, fostering progress in manipulating biomolecules, engineering cells, and computational biology. Agriculture and food production have significantly benefited from tools like high-throughput microarrays, accelerating the selection of desired traits. Genetic engineering, especially utilizing genome editing, facilitates precise alterations in plants and animals, harnessing microbiomes and fostering lab-grown meat production to alleviate environmental pressures. The emergence of new biotechnologies, notably genome editing, underscores the necessity for regulatory frameworks governing LM (living modified) organisms. Global regulations overseeing genetically engineered or genome-edited (GE) organisms, encompassing animals, exhibit considerable diversity. Nonetheless, prevailing international regulatory trends typically exclude genomeedited plants and animals, employing novel biotechnological techniques, from GMO/ LMO classification if they lack foreign genes and originate through natural mutations or traditional breeding programs. This comprehensive review scrutinizes ongoing risk and safety assessment cases, such as genome-edited beef cattle and fish in the USA and Japan. Furthermore, it investigates the limitations of existing regulations related to genome editing in Korea and evaluates newly proposed legislation, offering insights into the future trajectory of regulatory frameworks.

Analysis of Expressed Sequence Tags from the Red Alga Griffithsia okiensis

  • Lee, Hyoung-Seok;Lee, Hong-Kum;An, Gyn-Heung;Lee, Yoo-Kyung
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.541-546
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    • 2007
  • Red algae are distributed globally, and the group contains several commercially important species. Griffithsia okiensis is one of the most extensively studied red algal species. In this study, we conducted expressed sequence tag (ESTs) analysis and synonymous codon usage analysis using cultured G. okiensis samples. A total of 1,104 cDNA clones were sequenced using a cDNA library made from samples collected from Dolsan Island, on the southern coast of Korea. The clustering analysis of these sequences allowed for the identification of 1,048 unigene clusters consisting of 36 consensus and 1,012 singleton sequences. BLASTX searches generated 532 significant hits (E-value <$10^{-4}$) and via further Gene Ontology analysis, we constructed a functional classification of 434 unigenes. Our codon usage analysis showed that unigene clusters with more than three ESTs had higher GC contents (76.5%) at the third position of the codons than the singletons. Also, the majority of the optimal codons of G. okiensis and Chondrus crispus belonging to Bangiophycidae were G-ending, whereas those of Porphyra yezoensis belonging to Florideophycidae were G-ending. An orthologous gene search for the P. yezoensis EST database resulted in the identification of 39 unigenes commonly expressed in two rhodophytes, which have putative functions for structural proteins, protein degradation, signal transduction, stress response, and physiological processes. Although experiments have been conducted on a limited scale, this study provides a material basis for the development of microarrays useful for gene expression studies, as well as useful information for the comparative genomic analysis of red algae.

Development of a Reproducibility Index for cDNA Microarray Experiments

  • Kim, Byung-Soo;Rha, Sun-Young
    • Proceedings of the Korean Statistical Society Conference
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    • 2002.05a
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    • pp.79-83
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    • 2002
  • Since its introduction in 1995 by Schena et al. cDNA microarrays have been established as a potential tool for high-throughput analysis which allows the global monitoring of expression levels for thousands of genes simultaneously. One of the characteristics of the cDNA microarray data is that there is inherent noise even after the removal of systematic effects in the experiment. Therefore, replication is crucial to the microarray experiment. The assessment of reproducibility among replicates, however, has drawn little attention. Reproducibility may be assessed with several different endpoints along the process of data reduction of the microarray data. We define the reproducibility to be the degree with which replicate arrays duplicate each other. The aim of this note is to develop a novel measure of reproducibility among replicates in the cDNA microarray experiment based on the unprocessed data. Suppose we have p genes and n replicates in a microarray experiment. We first develop a measure of reproducibility between two replicates and generalize this concept for a measure of reproducibility of one replicate against the remaining n-1 replicates. We used the rank of the outcome variable and employed the concept of a measure of tracking in the blood pressure literature. We applied the reproducibility measure to two sets of microarray experiments in which one experiment was performed in a more homogeneous environment, resulting in validation of this novel method. The operational interpretation of this measure is clearer than Pearson's correlation coefficient which might be used as a crude measure of reproducibility of two replicates.

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Hypermethylation of the Ras Association Domain Family 1A (RASSF1A) Gene in Gallbladder Cancer

  • Kee, Se Kook;Lee, Ji Yun;Kim, Mi Jin;Lee, Su Man;Jung, Young Won;Kim, Young Joo;Park, Jae Yong;Bae, Han Ik;Hong, Hae Sook;Yun, Young Kook;Kim, Sang Geol;Kim, Dong Sun
    • Molecules and Cells
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    • v.24 no.3
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    • pp.364-371
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    • 2007
  • The tumor suppressor gene Ras association domain family 1A (RASSF1A) is highly methylated in a wide range of human sporadic tumors. The current study investigated the hypermethylation of RASSF1A, the expression of RASSF1A protein, and the correlation between these and the clinicopathological features of gallbladder (GB) cancer in Korean patients. Formalin-fixed, paraffin-embedded tumors and non-neoplastic GB tissues (22 carcinomas, 8 adenomas, 26 normal epithelia) were collected from patients who had undergone surgical resection. The methylation status of two regions of the RASSF1A CpG island was determined by methylation-specific PCR (MSP), and the expression of RASSF1A protein was examined by immunohistochemistry using tissue microarrays. The K-RAS mutation was analyzed by direct sequencing. Methylation of the RASSF1A promoter (region 1) was detected in 22.7% (5/22) of carcinomas, 12.5% (1/8) of adenomas, and 0% (0/26) of normal gallbladder epithelia (P = 0.025). Methylation of the first exon (region 2) was found in 36.4% (8/22) of carcinomas, 25.0% (2/8) of adenomas, and 8.0% (2/26) of normal gallbladder epithelia (P = 0.038). K-RAS mutations were present in 4.5% (1/22) of carcinomas and 25% (2/8) of adenomas. RASSF1A methylaton was not associated with clinicopathological factors or K-ras mutation. Reduction or loss of RASSF1A expression was observed in most methylated adenocarcinomas. Three RASSF1A-expressing human biliary tract cancer cell lines examined contained unmethylated promoters and exons 1. These results suggest that downregulation of RASSF1A expression by DNA hypermethylation may be involved in GB carcinogenesis.

Hypernetwork Classifiers for Microarray-Based miRNA Module Analysis (마이크로어레이 기반 miRNA 모듈 분석을 위한 하이퍼망 분류 기법)

  • Kim, Sun;Kim, Soo-Jin;Zhang, Byoung-Tak
    • Journal of KIISE:Software and Applications
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    • v.35 no.6
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    • pp.347-356
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    • 2008
  • High-throughput microarray is one of the most popular tools in molecular biology, and various computational methods have been developed for the microarray data analysis. While the computational methods easily extract significant features, it suffers from inferring modules of multiple co-regulated genes. Hypernetworhs are motivated by biological networks, which handle all elements based on their combinatorial processes. Hence, the hypernetworks can naturally analyze the biological effects of gene combinations. In this paper, we introduce a hypernetwork classifier for microRNA (miRNA) profile analysis based on microarray data. The hypernetwork classifier uses miRNA pairs as elements, and an evolutionary learning is performed to model the microarray profiles. miTNA modules are easily extracted from the hypernetworks, and users can directly evaluate if the miRNA modules are significant. For experimental results, the hypernetwork classifier showed 91.46% accuracy for miRNA expression profiles on multiple human canters, which outperformed other machine learning methods. The hypernetwork-based analysis showed that our approach could find biologically significant miRNA modules.

Effect of Extremely Low Frequency Magnetic Fields on Gene Expression in Human Mammary Epithelial MCF10A Cells

  • Hong, Mi-Na;Lee, Hyung-Chul;Kim, Bong Cho;Lee, Yun-Sil;Gimm, Yoon-Myung;Myung, Sung-Ho;Lee, Jae-Seon
    • Journal of electromagnetic engineering and science
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    • v.12 no.4
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    • pp.271-279
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    • 2012
  • The effects of extremely low frequency magnetic fields (ELF-MFs) on physiological processes at the cellular level remain unclear despite a number of studies. To investigate the effects of ELF-MFs on gene expression, we exposed human mammary epithelial MCF10A cells to fields of 1 mT magnetic flux density at 60 Hz for 4 and 16 h and measured the transcriptional responses of 24,000 genes using Illumina microarrays. In three independent experiments, we found no statistically significant alteration of expression levels for any of the genes assayed using a cutoff value of 1.2-fold. To confirm this result, we selected six genes with trends suggesting possible expression level changes, although these trends were not statistically significant, and investigated their expression levels further using a semiquantitative reverse-transcription polymerase chain reaction. In three independent experiments, we did not find any alterations in the expression levels of these genes. From these results, we conclude that ELF-MFs do not affect gene expression profiles under our exposure conditions.