• 제목/요약/키워드: Gene Targeting

검색결과 474건 처리시간 0.022초

Targeting Super-Enhancers for Disease Treatment and Diagnosis

  • Shin, Ha Youn
    • Molecules and Cells
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    • 제41권6호
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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.

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

  • 김영중;남경희;백인순;박정호;정순천;한지학;김창기
    • 농업과학연구
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    • 제41권3호
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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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    • 제43권2호
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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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    • 제40권1호
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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.

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

  • 임성렬;이진형;이효연;서석철
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2002년도 춘계 학술대회지
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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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    • 제7권3호
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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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    • 제11권1호
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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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    • 제27권7호
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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.

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

  • 정희;문정환;이성찬;유희주
    • 원예과학기술지
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    • 제29권5호
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    • pp.461-466
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    • 2011
  • 유전 육종 연구를 위해 연구자들은 실험 목적에 따라 다양한 종류의 프라이머를 제작해야 한다. 인터넷 상에서 다양한 공용 프로그램이 이용되고 있으나 많은 경우 사용자 편의성이 낮기 때문에 유전자의 구조를 고려하여 프라이머를 디자인하기 위해서는 시간과 노력이 소요된다. 본 연구에서는 엑손과 인트론 지역을 시각적으로 구별하면서 손쉽게 프라이머를 제작할 수 있는 프로그램인 Pickprimer를 개발하였다. 이 프로그램은 공용 프로그램인 Spidey와 Primer3 프로그램의 소스 코드를 결합한 후 그래픽 인터페이스를 추가하여 사용자가 유전자의 구조를 예측하고 이를 바탕으로 프라이머를 손쉽게 제작할 수 있게 했다. 입력 정보는 공용 데이터베이스에서 내려 받은 서열을 복사-붙임하여 이용할 수 있게 하였으며, 유전자의 구조를 그림으로 표현하고 동시에 엑손과 인트론 서열을 구별할 수 있게 했다. 이 프로그램을 이용하여 배추의 단일 카피 유전자에 대한 24 쌍의 프라이머를 디자인하고 6개 고정 품종을 대상으로 PCR과 전기영동 실험을 수행한 결과 제작한 모든 프라이머 쌍이 명확한 단일 밴드를 성공적으로 증폭시켰다. 이 프로그램은 분자표지의 개발뿐만 아니라 유전자 기능 연구 등 다양한 종류의 유전 육종 실험에 유용하게 이용될 수 있을 것으로 기대된다.

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

  • 정인실;방성호
    • 미생물학회지
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    • 제47권1호
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    • pp.1-6
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    • 2011
  • 암의 유전자치료에서 암세포로의 선택적 유전자전달 매체의 부족은 치료효과의 감소를 야기하는 문제이다. 본 연구에서는 난소암 유전자치료의 효율을 높이기 위한 목적으로 난소암세포로 선택적인 유전자전달을 하도록 개량된 아데노바이러스 벡터를 제조하고, 그 효율성을 난소암세포주를 이용하여 조사하였다. 난소암세포에 과다발현하는 분자인 ErbB receptor를 표적하도록 아데노바이러스 외피단백질 fiber에 ErbB receptor에 대한 ligand인 heregulin으로부터 유래한 펩티드를 부착하였다. 53개의 아미노산으로 구성된 외부 펩티드를 fiber에 부착하였을 때 바이러스 감염에 중요한 기능을 하는 fiber 삼량체 구조 형성에 영향을 미치지 않았다. Fiber를 조작한 개량 아데노바이러스는 야생형 fiber를 가진 1세대 아데노바이러스 벡터에 비해 선택적으로 난소암으로 유전자를 전달하는 비율이 증가하였다. 특히 항암제에 저항성을 가진 난소암세포주 OVCAR3에서 유전자전달 효율이 약 5배 증가되었다. 따라서 난소암의 유전자치료에서 개량된 아데노바이러스로 치료 유전자를 전달하면 치료의 효율성을 향상시킬 수 있을 것이다.