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Myostatin gene knockout mediated by Cas9-D10A nickase in chicken DF1 cells without off-target effect

  • Lee, Jeong Hyo;Kim, Si Won;Park, Tae Sub
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.5
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    • pp.743-748
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    • 2017
  • Objective: Based on rapid advancement of genetic modification techniques, genomic editing is expected to become the most efficient tool for improvement of economic traits in livestock as well as poultry. In this study, we examined and verified the nickase of mutated CRISPR-associated protein 9 (Cas9) to modulate the specific target gene in chicken DF1 cells. Methods: Chicken myostatin which inhibits muscle cell growth and differentiation during myogenesis was targeted to be deleted and mutated by the Cas9-D10A nickase. After co-transfection of the nickase expression vector with green fluorescent gene (GFP) gene and targeted multiplex guide RNAs (gRNAs), the GFP-positive cells were sorted out by fluorescence-activated cell sorting procedure. Results: Through the genotyping analysis of the knockout cells, the mutant induction efficiency was 100% in the targeted site. Number of the deleted nucleotides ranged from 2 to 39 nucleotide deletion. There was no phenotypic difference between regular cells and knockout cells. However, myostatin protein was not apparently detected in the knockout cells by Western blotting. Additionally, six off-target sites were predicted and analyzed but any non-specific mutation in the off-target sites was not observed. Conclusion: The knockout technical platform with the nickase and multiplex gRNAs can be efficiently and stablely applied to functional genomics study in poultry and finally adapted to generate the knockout poultry for agribio industry.

Comparative analysis of commonly used peak calling programs for ChIP-Seq analysis

  • Jeon, Hyeongrin;Lee, Hyunji;Kang, Byunghee;Jang, Insoon;Roh, Tae-Young
    • Genomics & Informatics
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    • v.18 no.4
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    • pp.42.1-42.9
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    • 2020
  • Chromatin immunoprecipitation coupled with high-throughput DNA sequencing (ChIP-Seq) is a powerful technology to profile the location of proteins of interest on a whole-genome scale. To identify the enrichment location of proteins, many programs and algorithms have been proposed. However, none of the commonly used peak calling programs could accurately explain the binding features of target proteins detected by ChIP-Seq. Here, publicly available data on 12 histone modifications, including H3K4ac/me1/me2/me3, H3K9ac/me3, H3K27ac/me3, H3K36me3, H3K56ac, and H3K79me1/me2, generated from a human embryonic stem cell line (H1), were profiled with five peak callers (CisGenome, MACS1, MACS2, PeakSeq, and SISSRs). The performance of the peak calling programs was compared in terms of reproducibility between replicates, examination of enriched regions to variable sequencing depths, the specificity-to-noise signal, and sensitivity of peak prediction. There were no major differences among peak callers when analyzing point source histone modifications. The peak calling results from histone modifications with low fidelity, such as H3K4ac, H3K56ac, and H3K79me1/me2, showed low performance in all parameters, which indicates that their peak positions might not be located accurately. Our comparative results could provide a helpful guide to choose a suitable peak calling program for specific histone modifications.

Multifactorial Traits of SARS-CoV-2 Cell Entry Related to Diverse Host Proteases and Proteins

  • You, Jaehwan;Seok, Jong Hyeon;Joo, Myungsoo;Bae, Joon-Yong;Kim, Jin Il;Park, Man-Seong;Kim, Kisoon
    • Biomolecules & Therapeutics
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    • v.29 no.3
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    • pp.249-262
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    • 2021
  • The most effective way to control newly emerging infectious disease, such as the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, is to strengthen preventative or therapeutic public health strategies before the infection spreads worldwide. However, global health systems remain at the early stages in anticipating effective therapeutics or vaccines to combat the SARS-CoV-2 pandemic. While maintaining social distance is the most crucial metric to avoid spreading the virus, symptomatic therapy given to patients on the clinical manifestations helps save lives. The molecular properties of SARS-CoV-2 infection have been quickly elucidated, paving the way to therapeutics, vaccine development, and other medical interventions. Despite this progress, the detailed biomolecular mechanism of SARS-CoV-2 infection remains elusive. Given virus invasion of cells is a determining factor for virulence, understanding the viral entry process can be a mainstay in controlling newly emerged viruses. Since viral entry is mediated by selective cellular proteases or proteins associated with receptors, identification and functional analysis of these proteins could provide a way to disrupt virus propagation. This review comprehensively discusses cellular machinery necessary for SARS-CoV-2 infection. Understanding multifactorial traits of the virus entry will provide a substantial guide to facilitate antiviral drug development.

Relapsed Acute Myeloid Leukemia Presenting as Multiple Breast Masses: A Case Report (유방의 다발성 결절로 발현한 급성 골수성 백혈병 재발의 건: 증례 보고)

  • Pamela Sung;Jong Yoon Lee;A Jung Chu
    • Journal of the Korean Society of Radiology
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    • v.84 no.2
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    • pp.454-459
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    • 2023
  • Hematologic malignancy of the breast is very rare. Here, we report a case of relapsed acute myeloid leukemia (AML) presenting as multiple breast masses. A 77-year-old female visited an outpatient clinic reporting palpable masses in both breasts. She had a medical history of AML, which showed complete remission after nine cycles of chemotherapy. On mammography and ultrasonography, there were multiple masses correlated with her palpable symptoms accompanied by enlarged lymph nodes. Core needle biopsy immunohistochemistry (IHC) results indicated AML and blastic plasmacytoid dendritic cell neoplasm. AML was confirmed using bone marrow biopsy. Although very rare, when a patient with a history of hematologic malignancy presents a palpable mass in the breast, clinicians should conduct proper tissue analysis, including IHC stating for leukemic markers, to guide appropriate diagnosis and treatment.

Differentiating Plasma Regions Through the non-Linear Relationship between the Band-gap and the Deposition-rate of a-Si Thin Films (a-Si 막의 Band-gap과 Deposition-rate간의 비선형 거동을 통한 플라즈마 영역의 경계 규명)

  • Park, Sung-Yul L.;Kim, Hee Won;Kim, Sang Duk;Kim, Jong Hwan;Kim, Bum Sung;Lee, Don Hee
    • 한국신재생에너지학회:학술대회논문집
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    • 2010.06a
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    • pp.72.1-72.1
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    • 2010
  • Thin film a-Si solar cells deposited by PECVD have many advantages compared to the traditional crystalline Si solar cells. They do not require expensive Si wafer, the process temperature is relatively low, possibility of scaling up for mass production, etc. In order to produce thin film solar cells, understanding the relationship between the material characteristics and deposition conditions is important. It has been reported by many groups that the band gap of the a-Si material and the deposition rate has an linear relationship, when RF power is used to control both. However, when the process pressure is changed in order to control the deposition rate and the band gap, a diversion from the well known linear relationship occurs. Here, we explain this diversion by the deposition condition crossing different plasma regions in the Paschen curve with a simple model. This model will become a guide to which condition a-Si thin films must be fabricated in order to get a high quality film.

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Dentinogenesis imperfecta type II: A case report with 17 years of follow-up

  • Gama, Francisco Jose Reis;Correa, Isabella Sousa;Valerio, Claudia Scigliano;Ferreira, Emanuelle de Fatima;Manzi, Flavio Ricardo
    • Imaging Science in Dentistry
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    • v.47 no.2
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    • pp.129-133
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    • 2017
  • Dentinogenesis imperfecta is a dominant autosomal hereditary disorder of dentin formation that affects the deciduous and permanent teeth. Its etiology is characterized by inadequate cell differentiation during odontogenesis. The clinical characteristics of dentinogenesis imperfecta are discolored teeth with a translucency that varies from gray to brown or amber. Radiographically, the teeth exhibit pulp obliteration, thin and short roots, bell-shaped crowns, and periapical bone rarefaction. The aim of this report was to present a case of dentinogenesis imperfecta type II that was followed up over a 17-year period. This report also presents scanning electron microscopy images of the enamel and dentin, showing that both were altered in the affected teeth. The disease characteristics and the treatments that were administered are reported in this study to guide dentists with respect to the need for early diagnosis and adequate follow-up to avoid major sequelae.

Structural Characteristic Analysis of an Ultra-Precision Machine for Machining Large-Surface Micro-Features (초정밀 대면적 미세 형상 가공기의 구조 특성 해석)

  • Kim, Seok-ll;Lee, Won-Jae
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.31 no.12
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    • pp.1173-1179
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    • 2007
  • In recent years, research to machine large-surface micro-features has become important because of the light guide panel of a large-scale liquid crystal display and the bipolar plate of a high-capacity proton exchange membrane fuel cell. In this study, in order to realize the systematic design technology and performance improvements of an ultra-precision machine for machining the large-surface micro-features, a structural characteristic analysis was performed using its virtual prototype. The prototype consisted of gantry-type frame, hydrostatic feed mechanisms, linear motors, brushless DC servo motor, counterbalance mechanism, and so on. The loop stiffness was estimated from the relative displacement between the tool post and C-axis table, which was caused by a cutting force. Especially, the causes of structural stiffness deterioration were identified through the structural deformation analysis of sub-models.

The Development of Outdoor Augmented Reality System by GPS (GPS를 이용한 옥외용 증강현실 시스템 개발)

  • Choi, T.J.;Jang, B.T.;Han, S.H.;Kim, J.K.;Hur, W.
    • Proceedings of the IEEK Conference
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    • 2000.06c
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    • pp.153-156
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    • 2000
  • In this paper, we developed an outdoor augmented reality system which has remote real scenes acquisition ability. The real scenes acquisition system consists of Image acquisition system, tracking system, wireless data transceiver and power supply. The tracking system consists of Tans Vector and RT-20 which measures a position and attitude of the CCD camera that attached to the remote control helicopter. Wireless data transceiver system is utilized for data transmission of remote system that of attitude, position information, and real scenes data that acquired by the CCD camera. Maximum propelling power of remote control helicopter is 15Kg, so we used 7.2V li-ion cell as a power supply for system minimize. As the results of experiment, the developing system presented application possibility of remote information acquisition system such as construction simulation & estimation, broadcasting, tour guide.

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The Comparison of Field Uniformity and SAR between TEM, Loop and Combined Elemented 16 Channel Transmit Arrays (TEM 형과 Loop 형의 전송소자를 이용한 16채널 3T Body 전송코일의 자장 균일도와 안전도 비교)

  • Ryu, Yeunchul;Kim, Young Beom
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.63 no.3
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    • pp.404-408
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    • 2014
  • In this research we evaluate the excitation homogeneity and SAR of three different Tx arrays after B1+ Shimming in the human body at 3T. Through the simulations, we compared the field properties that are standard deviation and mean value of transverse magnetization for 1) strip line TEM array, 2) rectangular loop array, 3) combined array of strip line element and rectangular loop and shown the utilities of B1+ shimming in human body model. After B1+ shimming, it is evaluated four different types of SAR for body mesh through whole body simulation; those are average, maximum 1-cell, maximum 1-gram, and maximum 10-gram SAR. It appears that in this particular comparison an array based on strip line elements can produce better homogeneity and lower SAR than an array of rectangular loops or an array of combined elements (strip line and loop). While many factors are considered in designing coils for production, it is hoped that methods and results like these will be used in the future to guide decisions and maximize benefit.

Reliability Assessment Criteria of Organic Light Emitting Diode(OLED) (유기 발광 다이오드의 신뢰성 평가기준)

  • Hong, Won-Sik;Song, Byeong-Suk;Jeong, Hai-Sung;Lim, Jae-Hak
    • Journal of Applied Reliability
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    • v.9 no.2
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    • pp.131-148
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    • 2009
  • An organic light emitting diode (OLED), also light emitting polymer (LEP) and organic electro luminescence (OEL), is any light emitting diode (LED) whose emissive electroluminescent layer is composed of a film of organic compounds. The layer usually contains a polymer substance that allows suitable organic compounds to be deposited. They are deposited in rows and columns onto a flat carrier by a simple "printing" process. The resulting matrix of pixels can emit light of different colors. Such systems can be used in television screens, computer displays, small, portable system screens such as cell phones and PDAs, advertising, information and indication. OLEDs can also be used in light sources for general space illumination, and large-area light-emitting elements. In this paper, we develop the general guide line of the accelerated life test for assuring B10 life of AMOLED(Active Matrix Organic Light Emitting Diode) and PMOLED(Passive Matrix Organic Light Emitting Diode) which are widely used for display monitor less than 115 mm.

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