• Title/Summary/Keyword: gene targeting

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Targeting Super-Enhancers for Disease Treatment and Diagnosis

  • Shin, Ha Youn
    • Molecules and Cells
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    • v.41 no.6
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    • pp.506-514
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    • 2018
  • The transcriptional regulation of genes determines the fate of animal cell differentiation and subsequent organ development. With the recent progress in genome-wide technologies, the genomic landscapes of enhancers have been broadly explored in mammalian genomes, which led to the discovery of novel specific subsets of enhancers, termed super-enhancers. Super-enhancers are large clusters of enhancers covering the long region of regulatory DNA and are densely occupied by transcription factors, active histone marks, and co-activators. Accumulating evidence points to the critical role that super-enhancers play in cell type-specific development and differentiation, as well as in the development of various diseases. Here, I provide a comprehensive description of the optimal approach for identifying functional units of super-enhancers and their unique chromatin features in normal development and in diseases, including cancers. I also review the recent updated knowledge on novel approaches of targeting super-enhancers for the treatment of specific diseases, such as small-molecule inhibitors and potential gene therapy. This review will provide perspectives on using super-enhancers as biomarkers to develop novel disease diagnostic tools and establish new directions in clinical therapeutic strategies.

Gene Flow from GM Cabbage to Non-GM Control (유전자변형 양배추로부터 비유전자변형 모본으로의 유전자 이동성)

  • Kim, Young-Joong;Nam, Kyong Hee;Pack, In Soon;Park, Jung-Ho;Jeong, Soon-Chun;Harn, Chee Hark;Kim, Chang-Gi
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.157-161
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    • 2014
  • Understanding the gene flow from genetically modified (GM) crops to conventional crops is important to prevent and mitigate seed contamination caused by pollen-mediated gene flow. We conducted a field test to investigate the gene flow from diamondback moth resistant GM cabbage (Brassica oleracea var. capitata) containing cry1Ac1 gene, to a non-GM control line AD126. GM and non-GM cabbage plants were cultivated in the field and pollinated using Bombus terrestris under the nets during the flowering periods. After seeds were collected from non-GM plants, hybrids between them and the GM cabbages were screened by multiplex PCR targeting cry1Ac1 gene. Out of 878 germinated seedlings, 168 hybrids were found and the average gene flow frequency was 19.7%. Because cabbage is mainly pollinated by insect pollinators, large-scale field tests are needed to study gene flow of GM cabbage.

Mouse phenogenomics, toolbox for functional annotation of human genome

  • Kim, Il-Yong;Shin, Jae-Hoon;Seong, Je-Kyung
    • BMB Reports
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    • v.43 no.2
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    • pp.79-90
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    • 2010
  • Mouse models are crucial for the functional annotation of human genome. Gene modification techniques including gene targeting and gene trap in mouse have provided powerful tools in the form of genetically engineered mice (GEM) for understanding the molecular pathogenesis of human diseases. Several international consortium and programs are under way to deliver mutations in every gene in mouse genome. The information from studying these GEM can be shared through international collaboration. However, there are many limitations in utility because not all human genes are knocked out in mouse and they are not yet phenotypically characterized by standardized ways which is required for sharing and evaluating data from GEM. The recent improvement in mouse genetics has now moved the bottleneck in mouse functional genomics from the production of GEM to the systematic mouse phenotype analysis of GEM. Enhanced, reproducible and comprehensive mouse phenotype analysis has thus emerged as a prerequisite for effectively engaging the phenotyping bottleneck. In this review, current information on systematic mouse phenotype analysis and an issue-oriented perspective will be provided.

A Restrictive Virus Tropism, Latency and Reactivation of Pseudorabies Virus Following Irreversible Deletion of Bsrl Restriction Site in the Thymidine-kinase Gene

  • Mohd Lila Mohd Azmi;Zeenathul, Nazariah-Allaudin;Abdel-Wahid Saeed Ali;Che Abdul Rahim Mohamed;Kamarudin, Awag-Isa
    • Journal of Microbiology
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    • v.40 no.1
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    • pp.1-10
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    • 2002
  • At the dose of 1000 p.f.u. per mouse,100% mortality occurred in mice inoculated with wild-type pseudorabies virus (PrV). In contrast, upon stable deletion of 10 bp nucleotides at the Bsrl site within the TK gene, PrV was rendered to be completely apathogenic. The deletion also caused the virus to be less capable of replicating in respiratory as well as in nervous system tissues. Although animals were exposed to high titers of TK-deleted PrVs, the virus failed to replicate to a high titer as compared to the pathogenic parental virus. In contrast to previous studies the deletion in the TK gene did not prevent the virus from establishing latency. Upon immunosuppression, the latent virus? however, reactivated but replicated at low titers. Interestingly, TK-deleted virus established latency and reactivation, that are occurred only in trigeminal ganglia and the cerebrums and no other tissues involved. Following reactivation, there was no indication of virus shedding in respiratory tissues as confirmed by virus isolation and polymerase chain reaction (PCR) technique targeting at the gB gene of PrV, The non-pathogenic virus with non-shedding characteristics, upon reactivation of the latent virus, would be the important feature of a live virus vaccine candidate.

Development Strategy for functional rice improved with human lactoferrin and enhancement of nutrient compounds (인체 모유 단백질 및 영양 성분 강화 고부가가치 기능성 쌀 생산 벼 품종 개발 전략)

  • Rhim Seong-Lyul;Lee Jin-Hyoung;Lee Hyo-Yeon;Suh Suk-Cheol
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2002.05a
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    • pp.48-50
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    • 2002
  • A strategy for development of a functional rice in proved with human lactoferrin and enhancement of nutrient compounds was planned. For the purposes we have cloned and characterized a human lactoferrin cDNA from human mammary gland cDNA library A endosperm storage vacuole targeting sequence and the cDNA fragment was linked to endosperm specific glutelin promoter. The fusion gene fragment was inserted into a binary vector containing MAR gene. In addition a new ${\beta}$-galactosidase gene from Bifidobacterium of human was used as a reporter gene in the vector system, Rice plants showing a high concentration of amino acids in the endosperm cells were developed by using a biochemical mutation and bred for the transformation with the binary vector system Finally we have established a transformation method for the rice endosperm cells.

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Population Analysis of Korean and Japanese Toxic Alexandrium catenella Using PCR Targeting the Area Downstream of the Chloroplast PsbA Gene

  • Kim Choong-Jae;Kim Chang-Hoon;Sako Yoshihiko
    • Fisheries and Aquatic Sciences
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    • v.7 no.3
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    • pp.130-135
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    • 2004
  • The marine dinoflagellate genus Alexandrium, which produces PSP toxins, has a global distribution. As human-assisted dispersal of the species has been suggested, it is important to develop molecular tools to trace the dispersal pathway. To screen population-specific DNA sequences that differentiate Korean and Japanese A. catenella, we targeted the area downstream of the chloroplast psbA gene using PCR with population-specific DNA primers followed by RFLP (restriction fragment length polymorphism) analysis and sequencing. The RFLP patterns of the PCR products divided Korean and Japanese A. catenella regional isolates into three types: Korean, Japanese, and type CMC3, isolated from Korea. We sequenced the PCR products, but found no similar gene in a homology search. The molecular phylogeny inferred from the sequences separated the Korean and Japanese A. catenella strains, as did the RFLP patterns. However, the Japanese isolates included two slightly different sequences (types J and K), while the Korean sequence was the same as the Japanese K type. In addition, a unique sequence was found in the Korean strains CMC2 and CMC3. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers designed from the type J sequence yielded PCR products for Japanese strains only, showing that the unknown gene can be used for a population analysis of Korean and Japanese A. catenella.

Genetic Variation of Korean Masu Salmon (Oncorhynchus masou) Populations Inferred from Mitochondrial DNA Sequence Analysis

  • Yoon, Moon-Geun;Jin, Hyung-Joo;Seong, Ki-Baek;Jin, Deuk-Hee
    • Fisheries and Aquatic Sciences
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    • v.11 no.1
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    • pp.36-40
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    • 2008
  • We analyzed the nucleotide sequences of about 500 bp of the mitochondrial NADH dehydrogenase subunit 3 (ND3) gene to estimate the genetic variation of Korean masu salmon (Oncorhynchus masou) populations. DNA samples were collected from 104 river-only specimens and 52 anadromous specimens from three hatcheries and one river. There are no records of artificial release into the river. We amplified the ND3 gene by polymerase chain reaction, targeting areas that included parts of the cytochrome oxidase III gene and the NADH dehydrogenase subunit 4L gene, and defined 14 haplotypes based on 12 variable nucleotide sites in the examined region. Among the haplotypes, ten were specific to river-only specimens within hatchery populations. Haplotype diversity of river-only populations in hatcheries was higher than that of anadromous and wild populations. Pairwise population $F_{ST}$ estimates and neighbor-joining tree analyses inferred that anadromous and river-only populations were distinct. These results suggest that sequence polymorphism in the ND3 region may be a useful marker for analyzing the genetic variation and population structure of masu salmon.

Anaerobic Ammonium-Oxidizing Bacteria in Cow Manure Composting

  • Wang, Tingting;Cheng, Lijun;Zhang, Wenhao;Xu, Xiuhong;Meng, Qingxin;Sun, Xuewei;Liu, Huajing;Li, Hongtao;Sun, Yu
    • Journal of Microbiology and Biotechnology
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    • v.27 no.7
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    • pp.1288-1299
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    • 2017
  • Composting is widely used to transform waste into valuable agricultural organic fertilizer. Anaerobic ammonium-oxidizing (anammox) bacteria play an important role in the global nitrogen cycle, but their role in composting remains poorly understood. In the present study, the community structure, diversity, and abundance of anammox bacteria were analyzed using cloning and sequencing methods by targeting the 16S rRNA gene and the hydrazine oxidase gene (hzo) in samples isolated from compost produced from cow manure and rice straw. A total of 25 operational taxonomic units were classified based on 16S rRNA gene clone libraries, and 14 operational taxonomic units were classified based on hzo gene clone libraries. The phylogenetic tree analysis of the 16S rRNA gene and deduced HZO protein sequences from the corresponding encoding genes indicated that the majority of the obtained clones were related to the known anammox bacteria Candidatus "Brocadia," Candidatus "Kuenenia," and Candidatus "Scalindua." The abundances of anammox bacteria were determined by quantitative PCR, and between $2.13{\times}10^5$ and $1.15{\times}10^6$ 16S rRNA gene copies per gram of compost were found. This study provides the first demonstration of the existence of anammox bacteria with limited diversity in cow manure composting.

Pickprimer: A Graphic User Interface Program for Primer Design on the Gene Target Region (픽프라이머 : 유전자 목표 구간 탐색 모듈을 포함한 프라이머 제작 그래픽 프로그램)

  • Chung, Hee;Mun, Jeong-Hwan;Lee, Seung-Chan;Yu, Hee-Ju
    • Horticultural Science & Technology
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    • v.29 no.5
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    • pp.461-466
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    • 2011
  • In genetic and molecular breeding studies of plants, researchers need to design various kinds of primers based on their research purposes. So far many kinds of web- or script-based non-commercial programs for primer design are available. Because most of them do not include user interface for multipurpose usage including gene structure prediction and direct target selection on sequences, it has been a laborious work to design primers targeting on the exon or intron regions of interesting genes. Here we report a primer designing graphic user interface program, Pickprimer, that includes gene structure prediction and primer design modules by combining source codes of the Spidey and Primer3 programs. This program provides simple graphic user interface to input sequences and design primers. Genomic sequence and mRNA or coding sequence of genes can be copy and pasted or input as fasta or text files. Based on alignment of the input sequences using the Spidey module, a putative gene structure is graphically visualized along with exon-intron sequences of color codes. Primer design can be easily performed by dragging mouse on the displayed sequences or input primer targeting position with desirable values of primers. The output of designed primers with detailed information is provided by the Primer3 module. PCR evaluation of 24 selected primer sets successfully amplified single amplicons from six Brassica rapa cultivars. The Pickprimer will be a convenient tool for genetic and molecular breeding studies of plants.

Development of the Gene Therapy Vector for Targeting Ovarian Cancer Cells through ErbB Receptors (ErbB 수용체를 이용한 난소암세포 표적 유전자치료 벡터의 개발)

  • Joung, In-Sil;Bang, Seong-Ho
    • Korean Journal of Microbiology
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    • v.47 no.1
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    • pp.1-6
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    • 2011
  • Inefficiency of in vivo gene transfer using currently available vectors reflects a major hurdle in cancer gene therapy. Both viral and non-viral approaches have been described to improve gene transfer efficiency but suffer from a number of limitations. Here we tested an adenovirus carrying the small peptide ligand derived from heregulin${\beta}$ EGF-like domain onto fiber, the adenoviral capsid protein, to deliver transgene to ovarian cancer cells which overexpress ErbB, the cognate receptors for heregulin. The attachement of 53 amino acids to fiber didn't affect on the fiber's trimer structure that is critical for the viral entry to cells. The fiber-modified adenovirus can mediate entry and expression of a ${\beta}$-galactosidase into cancer cells in an increased efficiency compared the unmodified adenovirus. Particularly, the gene transfer efficiency was improved up to 5 times in OVCAR3 cells, an ovarian cancer cell line. Such transduction systems hold promise for delivering genes to ErbB receptor overexpressing cancer cells, and could be used for future cancer gene therapy.