• BMB Reports
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• v.49 no.4
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• pp.201-207
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• 2016

The CRISPR-Cas system has emerged as a fascinating and important genome editing tool. It is now widely used in biology, biotechnology, and biomedical research in both academic and industrial settings. To improve the specificity and efficiency of Cas nucleases and to extend the applications of these systems for other areas of research, an understanding of their precise working mechanisms is crucial. In this review, we summarize current studies on the molecular structures and dynamic functions of type I and type II Cas nucleases, with a focus on target DNA searching and cleavage processes as revealed by single-molecule observations.

• YAKHAK HOEJI
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• v.37 no.5
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• pp.504-510
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• 1993

Human insulin gene is consisted of the polymorphic region with the repeating units, the regulatory sequence, the structural gene including the intervening sequence, and 3'-flanking region. The polymerase chain reaction, which amplifies the target DNA between two specific primers, has been performed for the amplification of human insulin gene and simple one-step cloning of it into Escherichia coli. Out of 1727 nuceotides compared, only 4 sites were variable: 5'-regulatory region(G2101$\rightarrow$AGG); IVS I(T2401$\rightarrow$A); Exon II(C2411 deletion); IVS II(A2740 dejection). The variations at the G2101 and T2401 were the same as those found in one American allele. The other two variations were observed only in the specific Korean allele. And, the enzyme digestion patterns among normal, insulin dependent diabetes mellitus, and non-insulin dependent diabetes mellitus were the same. On the other hand, PCR method showed the possibility of the quickaccess for the polymorphic region in terms of the restriction fragment length of polymorphism.

• Journal of Sensor Science and Technology
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• v.18 no.4
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• pp.251-262
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• 2009

Biosensors exploit the specific binding between recognition molecule on the biosensor surface and target molecule in analyte and are used in the detection of specific biomolecules such as protein, DNA, cell, virus, etc., with a view towards developing analytical devices. Recently, application field of biosensors have been expanding from diagnosis to biodefense because they can basically serve as high performance devices. This review describes the basic information of biosensors including definition, classification, and operational principle. Moreover, we introduce micro/nano technology-based biosensors with better detection performance than traditional method and their application examples.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1993.04a
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• pp.167-167
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• 1993

신물질의 면역독성 평가방법을 체계화하고 독성기작을 규명하고저 기존 면역독성 유발물질을 대상으로 in vivo 및 in vitro 실험을 수행하였다. 먼저 비장세포-간세포 공동배양 시스템을 대사를 요구하는 물질의 면역독성 평가에 in vitro system으로 사용하고저 DMN을 대상으로 공동배양 조건 및 배지에 대한 연구를 수행하였다. 비장세포원으로 BALB/c 마우스를 사용하였고, 간세포원으로 Spraque-Dawley 랫드를 사용하였다. DMN은 홀몬이 첨가된 배지에서 항체 형성반응을 크게 저하시켰으나 배지의 종류에 따른 DMW의 대사에는 큰 영향이 없었다. 또한 procarcinogen인 DMBA에 의한 면역독성은 DMBA의 대사물에 의해 유도되며 DNA가 primary target 임을 규명하였으며, 2-acetylaminofluorene (AAE)에 의한 면역 억제작용의 기작에서 인터루킨의 영향에 대해 연구하였다. Protein methylase inhibitor인 SF와 SIBA웨 면역억제 작용을 연구한 결과 lymphoproliferative response에서 protein methylase 가 직접 또는 간접적으로 관여함이 입증되었다.

• Journal of Microbiology and Biotechnology
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• v.15 no.2
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• pp.451-454
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• 2005

Introduction of foreign genes into brain cells, such as neurons and astrocytes, is a powerful approach to study the gene function and regulation in the neuroscience field. Calcium phosphate precipitates have been shown to cause cytotoxicity in some mammalian cells and brain cells, thus leading to low transfection efficiency. Here, we describe a retrovirus-mediated gene delivery method to transduce foreign genes into brain cells. In an attempt to achieve higher gene delivery efficiency in these cells, we made several changes to the original method, including (1) use of a new packaging cell line, Phoenix ampho cells, (2) transfection of pMX retroviral DNA, (3) inclusion of 25 mM chloroquine in the transduction, and (4) 3- 5 h incubation of retroviruses with target cells. The results showed that the modified protocol resulted in a range of 40- 60% gene delivery efficiency in neurons and astrocytes. Furthermore, these results suggest the potential of the retrovirus-mediated gene delivery protocol being modified and adapted for other transfection-refractory cell lines and primary cells.

• Journal of Korean Medicine Rehabilitation
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• v.30 no.1
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• pp.63-78
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• 2020

Objectives To review the epigenetic modifications involved in chronic pain and to improve individualized intervention for the chronic pain. Methods Focused literature review. Results Significant laboratory and clinical data support that epigenetic modifications have a potential role for development of chronic pain. Conclusions Epigenetic approach may identify mechanisms critical to the development of chronic pain after injury, and may provide new pathways and target mechanisms for future treatment and individualized medicine.

• Proceedings of the Korean Biophysical Society Conference
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• 1996.07a
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• pp.19-19
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• 1996

Cyclic AMP receptor protein (CRP) from E. Coli plays a key role in regulation of the expression of more than 20 genes of the bacterium. CRP binds in the presence of cAMP to a specific target site near the promoter of each gene under its regulation. CRP is a dimer (Mr～47,000) of two identical subunits. There are two binding domains in the CRP monomer, one for the binding of the cAMP and the other for the binding of specific DNA sequences. (omitted)

• Biomedical Science Letters
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• v.25 no.4
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• pp.372-380
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• 2019

Mice with specific modified genes are useful means of studying development and disease. The CRISPR/Cas9 system is a very powerful and effective tool for generating genetically modified mice in a simple and fast manner. To generate human IgG light kappa constant knock-in mice, we tested by microinjection of a mixture of Cas9 protein, single-guide RNA and target homologous recombinant donor DNA into zygotes. We found that the injection of 10 ng/μL of Cas9 protein and crRNA/tracrRNA, rather than single guide RNA, induced the production of knock-in mice more effectively. Thus, our study provides valuable information that will help to improve the production of knock-in mice and contribute the successful generation of humanized Ab-producing mice in Korea.

• BMB Reports
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• v.48 no.10
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• pp.589-594
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• 2015

The shadow enhancer of the short gastrulation (sog) gene directs its sequential expression in the neurogenic ectoderm and the ventral midline of the developing Drosophila embryo. Here, we characterize three unusual features of the shadow enhancer midline activity. First, the minimal regions for the two different enhancer activities exhibit high overlap within the shadow enhancer, meaning that one developmental enhancer possesses dual enhancer activities. Second, the midline enhancer activity relies on five Single-minded (Sim)-binding sites, two of which have not been found in any Sim target enhancers. Finally, two linked Dorsal (Dl)- and Zelda (Zld)-binding sites, critical for the neurogenic ectoderm enhancer activity, are also required for the midline enhancer activity. These results suggest that early activation by Dl and Zld may facilitate late activation via the noncanonical sites occupied by Sim. We discuss a model for Zld as a pioneer factor and speculate its role in midline enhancer activity.

• Molecules and Cells
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• v.40 no.3
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• pp.169-177
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• 2017

Tissue-specific transcription is critical for normal development, and abnormalities causing undesirable gene expression may lead to diseases such as cancer. Such highly organized transcription is controlled by enhancers with specific DNA sequences recognized by transcription factors. Enhancers are associated with chromatin modifications that are distinct epigenetic features in a tissue-specific manner. Recently, super-enhancers comprising enhancer clusters co-occupied by lineage-specific factors have been identified in diverse cell types such as adipocytes, hair follicle stem cells, and mammary epithelial cells. In addition, noncoding RNAs, named eRNAs, are synthesized at super-enhancer regions before their target genes are transcribed. Many functional studies revealed that super-enhancers and eRNAs are essential for the regulation of tissue-specific gene expression. In this review, we summarize recent findings concerning enhancer function in tissue-specific gene regulation and cancer development.