• Molecules and Cells
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• 제38권6호
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• pp.475-481
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• 2015

Programmable nucleases, which include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and RNA-guided engineered nucleases (RGENs) repurposed from the type II clustered, regularly interspaced short palindromic repeats (CRISPR)-CRISPR-associated protein 9 (Cas9) system are now widely used for genome editing in higher eukaryotic cells and whole organisms, revolutionising almost every discipline in biological research, medicine, and biotechnology. All of these nucleases, however, induce off-target mutations at sites homologous in sequence with on-target sites, limiting their utility in many applications including gene or cell therapy. In this review, we compare methods for detecting nuclease off-target mutations. We also review methods for profiling genome-wide off-target effects and discuss how to reduce or avoid off-target mutations.

• BMB Reports
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• 제49권3호
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• pp.135-136
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• 2016

Small interfering RNAs (siRNAs) have been developed to intentionally repress a specific gene expression by directing RNA-induced silencing complex (RISC), mimicking the endogenous gene silencer, microRNAs (miRNAs). Although siRNA is designed to be perfectly complementary to an intended target mRNA, it also suppresses hundreds of off-targets by the way that miRNAs recognize targets. Until now, there is no efficient way to avoid such off-target repression, although the mode of miRNA-like interaction has been proposed. Rationally based on the model called "transitional nucleation" which pre-requires base-pairs from position 2 to the pivot (position 6) with targets, we developed a simple chemical modification which completely eliminates miRNA-like off-target repression (0%), achieved by substituting a nucleotide in pivot with abasic spacers (dSpacer or C3 spacer), which potentially destabilize the transitional nucleation. Furthermore, by alleviating steric hindrance in the complex with Argonaute (Ago), abasic pivot substitution also preserves near-perfect on-target activity (∼80-100%). Abasic pivot substitution offers a general means of harnessing target specificity of siRNAs to experimental and clinical applications where misleading and deleterious phenotypes from off-target repression must be considered.

• BMB Reports
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• 제57권1호
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• pp.12-18
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• 2024

Due to the development of CRISPR technology, the era of effective editing of target genes has arrived. However, the off-target problem that occurs when recognizing target DNA due to the inherent nature of CRISPR components remains the biggest task to be overcome in the future. In this review, the principle of inducing such unintended off-target editing is analyzed from the structural aspect of CRISPR, and the methodology that has been developed to reduce off-target editing until now is summarized.

• Journal of Plant Biotechnology
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• 제47권3호
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• pp.185-193
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• 2020

최근, 유전자교정 작물의 상업화 승인 건수가 급속하게 늘어나고 있으며, 국내에서도 유전자교정 작물의 개발에 대한 집중적인 투자를 통하여 국제경쟁력을 높이기 위하여 노력하고 있다. 그러나 기존의 유전자변형작물의 상업화 과정에서 끊임없이 제기되어온 인체 및 환경에 대한 잠재적인 위해성 논란이 유전자교정 작물에 대하여서도 제기되고 있다. 특히, 비의도적 돌연변이(off-target)가 가장 큰 논란의 중심이 되고 있다. 따라서 본 리뷰는 식물이 내포하고 있는 장점인 배수체, 체세포 돌연변이 그리고 자연 상태에서 아그로박테리아의 T-DNA 단편의 수평 전이로 창출된 자연적인 유전자변형작물과 기존에 상업화가 승인된 유전자변형작물 이벤트들의 게놈 내 비의도적 돌연변이 사례 등을 검토한 결과 유전자교정 작물에서 나타나는 대부분의 비의도적 돌연변이는 인체 및 환경에 미칠 수 있는 위해성을 우려할 만한 수준이 아니라고 할 수 있었다. 이에, 유전자교정 작물의 안전성 평가를 위하여 새로운 규정을 제정할 필요가 없으며 기존의 유전자변형작물의 안전 관리규정을 일부 "용어의 정의" 등만 개정하여 적용하면 충분할 것으로 사료 되었다.

• Asian-Australasian Journal of Animal Sciences
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• 제33권2호
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• pp.360-372
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• 2020

Objective: Specific genomic sites can be recognized and permanently modified by genome editing. The discovery of endonucleases has advanced genome editing in pigs, attenuating xenograft rejection and cross-species disease transmission. However, off-target mutagenesis caused by these nucleases is a major barrier to putative clinical applications. Furthermore, off-target mutagenesis by genome editing has not yet been addressed in pigs. Methods: Here, we generated genetically inheritable α-1,3-galactosyltransferase (GGTA1) knockout Yucatan miniature pigs by combining transcription activator-like effector nuclease (TALEN) and nuclear transfer. For precise estimation of genomic mutations induced by TALEN in GGTA1 knockout pigs, we obtained the whole-genome sequence of the donor cells for use as an internal control genome. Results: In-depth whole-genome sequencing analysis demonstrated that TALEN-mediated GGTA1 knockout pigs had a comparable mutation rate to homologous recombination-treated pigs and wild-type strain controls. RNA sequencing analysis associated with genomic mutations revealed that TALEN-induced off-target mutations had no discernable effect on RNA transcript abundance. Conclusion: Therefore, TALEN appears to be a precise and safe tool for generating genomeedited pigs, and the TALEN-mediated GGTA1 knockout Yucatan miniature pigs produced in this study can serve as a safe and effective organ and tissue resource for clinical applications.

• International Journal of Aeronautical and Space Sciences
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• 제2권2호
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• pp.73-81
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• 2001

This paper describes the procedure to develop a robust estimator design method for a target tracker that accounts for both structured real parameter uncertainties and unknown inputs. Two robust design approaches are combined: the Mini-p-Norm. design method to consider real parameter uncertainties and the $H_{\infty}$ design technique for unknown disturbances and unknown inputs. Constant estimator gains are computed that guarantee the robust performance of the estimator in the presence of parameter variations in the target model and unknown inputs to the target. The new estimator has two design parameters. One design parameter allows the trade off between small estimator error variance and low sensitivity to unknown parameter variations. Another design parameter allows the trade off between the robustness to real parameter variations and the robustness to unknown inputs. This robust estimator design method was applied to the longitudinal motion tracking problem of a T-38 aircraft.

• The Journal of Asian Finance, Economics and Business
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• 제6권3호
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• pp.41-53
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• 2019

Building a target capital structure is one of the most important decisions in corporate financial management. The purpose of this article is to identify the determinants of capital structure and adjustment mechanism toward the target leverage. The partial adjustment model was applied on a sample of 306 non-financial companies listed on Vietnam stock exchange market during the period of 2008-2017. By the fixed effect model estimation method, the research results have discovered the factors of growth opportunities, firm size, tangible fixed assets and firm's unique characteristics have a positive effect on the target capital structure of enterprises. Besides, profitability and dividend payment have a negative effect on the target capital structure of enterprises. Accordingly, the research results show that the average adjustment speed toward target leverage of the firms is 90.03%. Research results also demonstrate firms have higher or lower debt ratio than the target debt ratio, capital surplus or capital deficit also have an impact on the adjustment rate toward the target capital structure. The research results are consistent with the Dynamic Trade-off Theory. From this result, this article has provided policy implications for non-financial companies listed on Vietnam's stock market in building a reasonable target capital structure according to operating timeline to maximize enterprise value.

• E2M - 전기 전자와 첨단 소재
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• 제8권3호
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• pp.285-290
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• 1995

High quality $Y_{1}$Ba$_{2}$Cu$_{3}$$O_{7-x}$ thin films have been fabricated by pulsed Nd:YAG laser deposition using an unusual 'off-axis' target-substrate geometry. Various properties of superconducting $Y_{1}$Ba$_{2}$Cu$_{3}$$O_{7-x}$ thin films have been studied systematically as a function of oxygen pressure during the deposition, in both 'on-axis' and the unusual 'off-axis' target substrate geometries. In the 'off-axis' geometry, one can completely eliminate the so-called 'laser droplets' from the film surface and thus obtain smooth high quality films. It is found that films with optimum structural and electrical properties are obtained at a lower oxygen pressure range during the 'off-axis' deposition when compared with that required in the 'on-axis' deposition geometry.

• 대한전기학회:학술대회논문집
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• 대한전기학회 1995년도 추계학술대회 논문집 학회본부
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• pp.361-363
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• 1995

High quality $Y_1Ba_2Cu_3O_{7-x}$ thin films have been fabricated by pulsed Nd:YAG laser deposition using an unusual 'off-axis' target-substrate geometry. Various properties of superconducting $Y_1Ba_2Cu_3O_{7-x}$ thin films have been studied systematically as a function of oxygen pressure during the deposition, in both 'on-axis' and the unusual 'off-axis' target substrate geometry. In the 'off-axis' geometry, one can completely eliminate the so-called 'laser droplets' form the thin surface and thus obtain smooth high qualify films. It is found that films with optimum structural and electrical properties are obtained at a lower oxygen pressure range during the 'off-axis' deposition when compared with that required in the 'on-axis' deposition geometry.

• 제어로봇시스템학회논문지
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• 제11권7호
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• pp.587-594
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• 2005

In this paper, a robust correlation-based video tracking is proposed to track a moving object in correlated image sequences. A correlation-based video tracking algorithm seeks to align the incoming target image with the reference target block image, but has critical problems, so called a false-peak problem and a drift phenomenon (correlator walk-off. The false-peak problem is generally caused by highly correlated background pixels with similar intensity of a moving target and the drift phenomenon occurs when tracking errors accumulate from frame to frame because of the nature of the correlation process. At first, the false-peaks problem for the ordinary correlation-based video tracking is investigated using a simple mathematical analysis. And, we will suggest a robust selective-attention correlation measure with a gradient preprocessor combined by a drift removal compensator to overcome the walk-off problem. The drift compensator adaptively controls the template block size according to the target size of interest. The robustness of the proposed method for practical application is demonstrated by simulating two real-image sequences.