• Journal of Animal Reproduction and Biotechnology
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• v.34 no.3
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• pp.148-156
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• 2019

The Transgenic livestock can be useful for the production of disease-resistant animals, pigs for xenotranplantation, animal bioreactor for therapeutic recombinant proteins and disease model animals. Previously, conventional methods without using artificial nuclease-dependent DNA cleavage system were used to produce such transgenic livestock, but their efficiency is known to be low. In the last decade, the development of artificial nucleases such as zinc-finger necleases (ZFNs), transcription activator-like effector nucleases (TALENs) and clustered regulatory interspaced short palindromic repeat (CRISPR)/Cas has led to more efficient production of knock-out and knock-in transgenic livestock. However, production of knock-in livestock is poor. In mouse, genetically modified mice are produced by coinjecting a pair of knock-in vector, which is a donor DNA, with a artificial nuclease in a pronuclear fertilized egg, but not in livestock. Gene targeting efficiency has been increased with the use of artificial nucleases, but the knock-in efficiency is still low in livestock. In many research now, somatic cell nuclear transfer (SCNT) methods used after selection of cell transfected with artificial nuclease for production of transgenic livestock. In particular, it is necessary to develop a system capable of producing transgenic livestock more efficiently by co-injection of artificial nuclease and knock-in vectors into fertilized eggs.

• The Mathematical Education
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• v.62 no.3
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• pp.401-416
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• 2023

This study systematically examined the influence of preservice teachers' self-efficacy and AI anxiety, on the intention to use AI programs 'knock-knock! mathematics expedition' in mathematics lessons based on a technology acceptance model. The research model was established with variables including self-efficacy, AI anxiety, perceived ease of use, perceived usefulness, and intention of use from 254 pre-service teachers. The structural relationships and direct and indirect effects between these variables were examined through structural equation modeling. The results indicated that self-efficacy significantly affected perceived ease of use, perceived usefulness, and intention to use. In contrast, AI anxiety did not significantly influence perceived ease of use and perceived usefulness. Perceived ease of use significantly affected perceived usefulness and intention to use and perceived usefulness significantly affected intention to use. The findings offer insights and strategies for encouraging the use of 'knock-knock! mathematics expedition' by preservice teachers in mathematics lessons.

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.3
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• pp.99-108
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• 2023

Background: Efficient gene editing technology is needed for successful knock-in. Homologous recombination (HR) is a major double-strand break repair pathway that can be utilized for accurately inserting foreign genes into the genome. HR occurs during the S/G2 phase, and the DNA mismatch repair (MMR) pathway is inextricably linked to HR to maintain HR fidelity. This study was conducted to investigate the effect of inhibiting MMR-related genes using CdCl₂, an MMR-related gene inhibitor, on HR efficiency in HC11 cells. Methods: The mRNA and protein expression levels of MMR-related genes (Msh2, Msh3, Msh6, Mlh1, Pms2), the HR-related gene Rad51, and the NHEJ-related gene DNA Ligase IV were assessed in HC11 cells treated with 10 μM of CdCl₂ for 48 hours. In addition, HC11 cells were transfected with a CRISPR/sgRNA expression vector and a knock-in vector targeting Exon3 of the mouse-beta casein locus, and treated with 10 μM cadmium for 48 hours. The knock-in efficiency was monitored through PCR. Results: The treatment of HC11 cells with a high-dose of CdCl₂ decreased the mRNA expression of the HR-related gene Rad51 in HC11 cells. In addition, the inhibition of MMR-related genes through CdCl₂ treatment did not lead to an increase in knock-in efficiency. Conclusions: The inhibition of MMR-related gene expression through high-dose CdCl₂ treatment reduces the expression of the HR-related gene Rad51, which is active during recombination. Therefore, it was determined that CdCl₂ is an inappropriate compound for improving HR efficiency.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.11
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• pp.1644-1651
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• 2014

Transgenic animals have become important tools for the production of therapeutic proteins in the domestic animal. Production efficiencies of transgenic animals by conventional methods as microinjection and retrovirus vector methods are low, and the foreign gene expression levels are also low because of their random integration in the host genome. In this study, we investigated the homologous recombination on the porcine ${\beta}$-casein gene locus using a knock-in vector for the ${\beta}$-casein gene locus. We developed the knock-in vector on the porcine ${\beta}$-casein gene locus and isolated knock-in fibroblast for nuclear transfer. The knock-in vector consisted of the neomycin resistance gene (neo) as a positive selectable marker gene, diphtheria toxin-A gene as negative selection marker, and 5' arm and 3' arm from the porcine ${\beta}$-casein gene. The secretion of enhanced green fluorescent protein (EGFP) was more easily detected in the cell culture media than it was by western blot analysis of cell extract of the HC11 mouse mammary epithelial cells transfected with EGFP knock-in vector. These results indicated that a knock-in system using ${\beta}$-casein gene induced high expression of transgene by the gene regulatory sequence of endogenous ${\beta}$-casein gene. These fibroblasts may be used to produce transgenic pigs for the production of therapeutic proteins via the mammary glands.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2005.07a
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• pp.49-49
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• 2005

• Transactions of the Korean Society of Automotive Engineers
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• v.14 no.2
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• pp.84-91
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• 2006

A heavy duty hydrogen-natural gas fueled engine can obtain stable operation at ultra lean conditions and reduce emissions extremely. Reduction of $CO_2$ in its engine is one of the most benefit. In this study, rate of hydrogen addition($R_{dH2}$) and compression ratio($\varepsilon$) were investigated including performance of this engine. As results, it was found that phenomenon of pressure oscillation when increasing $R_{dH2}$ and $\varepsilon$, it means occurring knock. It consider that pressure oscillation was increased due to fast burning speed of hydrogen. Even if same compression ratio, pressure oscillation was remarkable increased according to increasing $R_{dH2}$. Therefore, limit compression ratio of knock occurring was reduced by increasing $R_{dH2}$.

• BMB Reports
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• v.56 no.2
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• pp.102-107
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• 2023

Genome editing using CRISPR-associated technology is widely used to modify the genomes rapidly and efficiently on specific DNA double-strand breaks (DSBs) induced by Cas9 endonuclease. However, despite swift advance in Cas9 engineering, structural basis of Cas9-recognition and cleavage complex remains unclear. Proper assembly of this complex correlates to effective Cas9 activity, leading to high efficacy of genome editing events. Here, we develop a CRISPR/Cas9-RAD51 plasmid constitutively expressing RAD51, which can bind to single-stranded DNA for DSB repair. We show that the efficiency of CRISPR-mediated genome editing can be significantly improved by expressing RAD51, responsible for DSB repair via homologous recombination (HR), in both gene knock-out and knock-in processes. In cells with CRISPR/Cas9-RAD51 plasmid, expression of the target genes (cohesin SMC3 and GAPDH) was reduced by more than 1.9-fold compared to the CRISPR/Cas9 plasmid for knock-out of genes. Furthermore, CRISPR/Cas9-RAD51 enhanced the knock-in efficiency of DsRed donor DNA. Thus, the CRISPR/Cas9-RAD51 system is useful for applications requiring precise and efficient genome edits not accessible to HR-deficient cell genome editing and for developing CRISPR/Cas9-mediated knockout technology.

• Journal of The Korean Society of Integrative Medicine
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• v.8 no.3
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• pp.1-10
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• 2020

Purpose : The purpose of this study is to analyze the corrective effect of the general coordinative manipulation (GCM) joint intervention model on distort leg with imbalance of the lower extremity joints, pelvic and shoulder girdles, and lumbar spine. Methods : The study used a comparative analysis of the size of the distort leg and the imbalance of the lower extremity joints, pelvic and shoulder girdles, and lumbar spine before and after the application of the GCM joint intervention model. A total of 31 subjects from movement center G and the department of physical therapy at university M were selected as research subjects, and they were divided into two groups. The GCM joint intervention model was applied to 18 subjects in the bow knee group and 13 subjects in the knock knee group. The two groups received daily intervention three times a week for four weeks. The corrective effect of the GCM joint intervention model for each type of distort leg was compared and analyzed. Results : The effects of the GCM joint intervention model in correcting bow knee and knock knee with knee deformation and imbalance of the lower extremity joints, pelvic and shoulder girdles, and lumbar spine were significant in most domains (p<.05). The correlation between the bow knee and knock knee groups showed significance in most domains (p<.05). Conclusion : The GCM joint intervention model showed significant corrective effect in the bow knee and knock knee groups in terms of knee deformation, lower extremity joints, pelvic and shoulder girdles, and lumbar spine (p<.05).

• Nuclear Engineering and Technology
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• v.48 no.2
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• pp.482-494
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• 2016

The Feshbache-Kermane-Koonin (FKK) multi-step direct (MSD) theory of pre-equilibrium reactions has been used to compute the single-step cross-sections for some (p,${\alpha}$) reactions using the knock-on and pick-up reaction mechanisms at two incident proton energies. For the knock-on mechanism, the reaction was assumed to have taken place by the direct ejection of a preformed alpha cluster in a shell-model state of the target. But the reaction was assumed to have taken place by the pick-up of a preformed triton cluster (also bound in a shell-model state of the target core) by the incident proton for the pick-up mechanism. The Yukawa forms of potential were used for the proton-alpha (for the knock-on process) and proton-triton (for the pick-up process) interaction and several parameter sets for the proton and alpha-particle optical potentials. The calculated cross-sections for both mechanisms gave satisfactory fits to the experimental data. Furthermore, it has been shown that some combinations of the calculated distorted wave Born approximation cross-sections for the two reaction mechanisms in the FKK MSD theory are able to give better fits to the experimental data, especially in terms of range of agreement. In addition, the theory has been observed to be valid over a wider range of energy.

• The Journal of Natural Sciences
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• v.15 no.1
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• pp.57-67
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• 2005

In Order to investigate the physiological role of thiol peroxidase in Bacillus subtilis, a thiol peroxidase (btpx) knock-out mutant was generated by homologous recombination. The growth of btpx knock-out mutant in aerobic condition showed a similar pattern with that of wild type of Bacillus subtilis 168/ But btpx knock-out mutant showed a retarded growht in response to oxidative stress such as $H_2O_2$, cumene hydroperoxide (CHP) treatments.