• Title/Summary/Keyword: Genetic Approach

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Utilizing cell-free DNA to validate targeted disruption of MYO7A in rhesus macaque pre-implantation embryos

  • Junghyun Ryu;Fernanda C. Burch;Emily Mishler;Martha Neuringer;Jon D. Hennebold;Carol Hanna
    • Journal of Animal Reproduction and Biotechnology
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    • v.37 no.4
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    • pp.292-297
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    • 2022
  • Direct injection of CRISPR/Cas9 into zygotes enables the production of genetically modified nonhuman primates (NHPs) essential for modeling specific human diseases, such as Usher syndrome, and for developing novel therapeutic strategies. Usher syndrome is a rare genetic disease that causes loss of hearing, retinal degeneration, and problems with balance, and is attributed to a mutation in MYO7A, a gene that encodes an uncommon myosin motor protein expressed in the inner ear and retinal photoreceptors. To produce an Usher syndrome type 1B (USH1B) rhesus macaque model, we disrupted the MYO7A gene in developing zygotes. Identification of appropriately edited MYO7A embryos for knockout embryo transfer requires sequence analysis of material recovered from a trophectoderm (TE) cell biopsy. However, the TE biopsy procedure is labor intensive and could adversely impact embryo development. Recent studies have reported using cell-free DNA (cfDNA) from embryo culture media to detect aneuploid embryos in human in vitro fertilization (IVF) clinics. The cfDNA is released from the embryo during cell division or cell death, suggesting that cfDNA may be a viable resource for sequence analysis. Moreover, cfDNA collection is not invasive to the embryo and does not require special tools or expertise. We hypothesized that selection of appropriate edited embryos could be performed by analyzing cfDNA for MYO7A editing in embryo culture medium, and that this method would be advantageous for the subsequent generation of genetically modified NHPs. The purpose of this experiment is to determine whether cfDNA can be used to identify the target gene mutation of CRISPR/Cas9 injected embryos. In this study, we were able to obtain and utilize cfDNA to confirm the mutagenesis of MYO7A, but the method will require further optimization to obtain better accuracy before it can replace the TE biopsy approach.

Prediction of Germination of Korean Red Pine (Pinus densiflora) Seed using FT NIR Spectroscopy and Binary Classification Machine Learning Methods (FT NIR 분광법 및 이진분류 머신러닝 방법을 이용한 소나무 종자 발아 예측)

  • Yong-Yul Kim;Ja-Jung Ku;Da-Eun Gu;Sim-Hee Han;Kyu-Suk Kang
    • Journal of Korean Society of Forest Science
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    • v.112 no.2
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    • pp.145-156
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    • 2023
  • In this study, Fourier-transform near-infrared (FT-NIR) spectra of Korean red pine seeds stored at -18℃ and 4℃ for 18 years were analyzed. To develop seed-germination prediction models, the performance of seven machine learning methods, namely XGBoost, Boosted Tree, Bootstrap Forest, Neural Networks, Decision Tree, Support Vector Machine, PLS-DA, were compared. The predictive performance, assessed by accuracy, misclassification, and area under the curve (0.9722, 0.0278, and 0.9735 for XGBoost, and 0.9653, 0.0347, and 0.9647 for Boosted Tree), was better for the XGBoost and decision tree models when compared with other models. The 54 wave-number variables of the two models were of high relative importance in seed-germination prediction and were grouped into six spectral ranges (811~1,088 nm, 1,137~1,273 nm, 1,336~1,453 nm, 1,666~1,671 nm, 1,879~2,045 nm, and 2,058~2,409 nm) for aromatic amino acids, cellulose, lignin, starch, fatty acids, and moisture, respectively. Use of the NIR spectral data and two machine learning models developed in this study gave >96% accuracy for the prediction of pine-seed germination after long-term storage, indicating this approach could be useful for non-destructive viability testing of stored seed genetic resources.

Availability of the metapopulation theory in research of biological invasion: Focusing on the invasion success (침입생물 연구에 대한 메타개체군 이론의 활용 가능성: 침입 성공을 중심으로)

  • Jaejun Song;Jinsol Hong;Kijong Cho
    • Korean Journal of Environmental Biology
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    • v.40 no.4
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    • pp.525-549
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    • 2022
  • The process of biological invasion is led by the dynamics of a population as a demographic and evolutionary unit. Spatial structure can affect the population dynamics, and it is worth being considered in research on biological invasion which is always accompanied by dispersal. Metapopulation theory is a representative approach to spatially structured populations, which is chiefly applied in the field of ecology and evolutionary biology despite the controversy about its definition. In this study, metapopulation was considered as a spatially structured population that includes at least one subpopulation with significant extinction probability. The early phase of the invasion is suitable to be analyzed in aspects of the metapopulation concept because the introduced population usually has a high extinction probability, and their ecological·genetic traits determining the invasiveness can be affected by the metapopulation structure. Although it is important in the explanation of the prediction of the invasion probability, the metapopulation concept is rarely used in ecological research about biological invasion in Korea. It is expected that applying the metapopulation theory can supply a more detailed investigation of the invasion process at the population level, which is relatively inadequate in Korea. In this study, a framework dividing the invasive metapopulation into long- and middle-distance scales by the relative distance of movement to the natural dispersal range of species is proposed to easily analyze the effect of a metapopulation in real cases. Increased understanding of the mechanisms underlying invasions and improved prediction of future invasion risk are expected with the metapopulation concept and this framework.

Epistatic Interaction Analysis of Two Dull Genes, wx-mq and du1, Affecting Amylose Content Using Nearly Isogenic Lines in Rice

  • Ju-Won Kang;Ji-Yoon Lee;Gi-Un Seong;Youngho Kwon;So-Myeong Lee;Dong Jin Shin;Sais-Beul Lee;Hyunnggon Mang;Dong Soo Park;Jong-Hee Lee;Jun-Hyeon Cho;Gi-Won Oh
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.267-267
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    • 2022
  • Glutinous rice is a key grain quality trait occupying an important part during rice processing in most rice growing areas. Amylose content (AC) of rice determine eating quality which is one of the major traits in rice breeding program. In this study, a gene pyramiding approach was used to introduce two dull genes, responsible for low amylose contents, for glutinous rice breeding using marker assisted selection (MAS). Two dull genes were located on chromosome 6 (wx-mq, AC: 12.7 %) and chromosome 10 (du1, AC: 10.3%), respectively. To test whether these two dull genes have an epistatic interaction, we developed an F2 population by crossing two nearly isogenic lines(NILs) harboring wx-mq and du1. Gene based marker and KASP marker were used to select NILs(NIL-nor, NIL-wxmq, NIL-du1, and NIL-wxmq/du1) from the F2 population. A two-way ANOVA revealed an epistatic interaction between the two genes in the F2 population. The mean of Amylose contents for NIL-nor, NIL-wxwq, NIL-(du1, and NIL-wxmq/du1 were 17.3%, 12.5%, 9.7%, and 7.2%, respectively. This interaction was confirmed by an analysis of NILs indicating that both genes are involved in the same genetic mechanism controlling amylose contents. This result will be useful for rice breeding related to amylose content.

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(Image Analysis of Electrophoresis Gels by using Region Growing with Multiple Peaks) (다중 피크의 영역 성장 기법에 의한 전기영동 젤의 영상 분석)

  • 김영원;전병환
    • Journal of KIISE:Software and Applications
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    • v.30 no.5_6
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    • pp.444-453
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    • 2003
  • Recently, a great interest of bio-technology(BT) is concentrated and the image analysis technique for electrophoresis gels is highly requested to analyze genetic information or to look for some new bio-activation materials. For this purpose, the location and quantity of each band in a lane should be measured. In most of existing techniques, the approach of peak searching in a profile of a lane is used. But this peak is improper as the representative of a band, because its location does not correspond to that of the brightest pixel or the center of gravity. Also, it is improper to measure band quantity in most of these approaches because various enhancement processes are commonly applied to original images to extract peaks easily. In this paper, we adopt an approach to measure accumulated brightness as a band quantity in each band region, which Is extracted by not using any process of changing relative brightness, and the gravity center of the region is calculated as a band location. Actually, we first extract lanes with an entropy-based threshold calculated on a gel-image histogram. And then, three other methods are proposed and applied to extract bands. In the MER method, peaks and valleys are searched on a vertical search line by which each lane is bisected. And the minimum enclosing rectangle of each band is set between successive two valleys. On the other hand, in the RG-1 method, each band is extracted by using region growing with a peak as a seed, separating overlapped neighbor bands. In the RG-2 method, peaks and valleys are searched on two vertical lines by which each lane is trisected, and the left and right peaks nay be paired up if they seem to belong to the same band, and then each band region is grown up with a peak or both peaks if exist. To compare above three methods, we have measured the location and amount of bands. As a result, the average errors in band location of MER, RG-1, and RG-2 were 6%, 3%, and 1%, respectively, when the lane length is normalized to a unit value. And the average errors in band amount were 8%, 5%, and 2%, respectively, when the sum of band amount is normalized to a unit value. In conclusion, RG-2 was shown to be more reliable in the accuracy of measuring the location and amount of bands.

Autopoietic Machinery and the Emergence of Third-Order Cybernetics (자기생산 기계 시스템과 3차 사이버네틱스의 등장)

  • Lee, Sungbum
    • Cross-Cultural Studies
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    • v.52
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    • pp.277-312
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    • 2018
  • First-order cybernetics during the 1940s and 1950s aimed for control of an observed system, while second-order cybernetics during the mid-1970s aspired to address the mechanism of an observing system. The former pursues an objective, subjectless, approach to a system, whereas the latter prefers a subjective, personal approach to a system. Second-order observation must be noted since a human observer is a living system that has its unique cognition. Maturana and Varela place the autopoiesis of this biological system at the core of second-order cybernetics. They contend that an autpoietic system maintains, transforms and produces itself. Technoscientific recreation of biological autopoiesis opens up to a new step in cybernetics: what I describe as third-order cybernetics. The formation of technoscientific autopoiesis overlaps with the Fourth Industrial Revolution or what Erik Brynjolfsson and Andrew McAfee call the Second Machine Age. It leads to a radical shift from human centrism to posthumanity whereby humanity is mechanized, and machinery is biologized. In two versions of the novel Demon Seed, American novelist Dean Koontz explores the significance of technoscientific autopoiesis. The 1973 version dramatizes two kinds of observers: the technophobic human observer and the technology-friendly machine observer Proteus. As the story concludes, the former dominates the latter with the result that an anthropocentric position still works. The 1997 version, however, reveals the victory of the techno-friendly narrator Proteus over the anthropocentric narrator. Losing his narrational position, the technophobic human narrator of the story disappears. In the 1997 version, Proteus becomes the subject of desire in luring divorcee Susan. He longs to flaunt his male egomaniac. His achievement of male identity is a sign of technological autopoiesis characteristic of third-order cybernetics. To display self-producing capabilities integral to the autonomy of machinery, Koontz's novel demonstrates that Proteus manipulates Susan's egg to produce a human-machine mixture. Koontz's demon child, problematically enough, implicates the future of eugenics in an era of technological autopoiesis. Proteus creates a crossbreed of humanity and machinery to engineer a perfect body and mind. He fixes incurable or intractable diseases through genetic modifications. Proteus transfers a vast amount of digital information to his offspring's brain, which enables the demon child to achieve state-of-the-art intelligence. His technological editing of human genes and consciousness leads to digital standardization through unanimous spread of the best qualities of humanity. He gathers distinguished human genes and mental status much like collecting luxury brands. Accordingly, Proteus's child-making project ultimately moves towards technologically-controlled eugenics. Pointedly, it disturbs the classical ideal of liberal humanism celebrating a human being as the master of his or her nature.

Generation of a transgenic mouse model to study cranial suture development; Apert syndrome (두개봉합 발육 연구를 위한 형질변환 쥐의 개발 : 어퍼트 신드롬)

  • Lee, Kee-Joon;Ratisoontorn, Chootima;Baik, Hyoung-Seon;Park, Young-Chel;Park, Kwang-Kyun;Nah, Hyun-Duck
    • The korean journal of orthodontics
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    • v.33 no.6 s.101
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    • pp.485-497
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    • 2003
  • The form and function of the craniofacial structure critically depend on genetic information. With recent advances in the molecular technology, genes that are important for normal growth and morphogenesis of the craniofacial skeleton are being rapidly uncovered, shaping up modem craniofacial biology. One of them is fibroblast growth factor receptor 2 (FGFR2). Specific point mutations in the. FGFR2 gene have been linked to Apert syndrome, which is characterized by premature closure of cranial sutures and craniofacial anomalies as well as limb deformities. To study pathogenic mechanisms underlying craniosynostosis phenotype of Apert syndrome, we used a transgenic approach; an FGFR2 minigene construct containing an Apert mutation (a point mutation that substitute proline at the position 253 to arginine; P253R) was introduced into fertilized mouse germ cells by DNA microinjection. The injected cells were then allowed to develop into transgenic mice. We used a bone-specific promoter (a DNA fragment from the type I collagen gene) to confine the expression of mutant FGFR2 gene to the bone tissue, and asked whether expression of mutant FGFR2 in bone is sufficient to cause the craniosynostosis phenotype in mice. Initial characterization of these mice shows prematurely closed cranial sutures with facial deformities expected from Apert patients. We also demonstrate that the transgene produces mutant FGFR2 protein with increased functional activities. Having this useful mouse model, we now can ask questions regarding the role of FGFR2 in normal and abnormal development of cranial bones and sutures.

The review on the identification of the gifted child in the sports (스포츠영재의 판별에 관한 고찰)

  • 신승윤
    • Journal of Gifted/Talented Education
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    • v.11 no.1
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    • pp.81-98
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    • 2001
  • The purpose of this study was to indicate the ways to identify the gifted child in the sports, and the research direction in this area in the future. The most of researches related to the early development of the children who showed the gifted talents in the certain sports skills were published from the European area such as the old East Germany or Soviet Union. Those research papers or articles were main focus of this study to be analyzed. The analysis in this study will be assistant to provide the direction of the research in the future about the method to identify appropriately on the children with the gifted sports skills. -Related to the period of selection of the children with sports skill gifted, the divided selection procedure will be good such as the 1st selection and 2nd selection. In the 1st selection, the goal will be to test the potentials of the children with the sports talents in the public school with creating the special group, and the 2nd selection will be to examine the qualification of the admission to the special schools for the children with the sports talents. -Related to the method of selection of the children with sports talents, the special item on the developmental procedure should be considered on the each field as well as the potentials focused. And, the physical factors, the condition of growth and development, psychological factor, and environmental factor should be comprehensively considered. -The system of promotion in the period of the primary school will be appropriate in the special group for the children with the sports talents, and the special school will be good from the middle school. The system outlooks will be adequate if shaped as the hierarchical structure. -The curriculum for the children with the sports talents will be proper that the basic fitness for the earlier ages and the professional skills will be recommended if getting older. -The continuous research is strongly required for the judgement and promotion of the children with sports talents, therefore, the multiple intelligences approach will be helpful for the development of the bodily-kinesthetic intelligence, and in other words, the new era of the genetic method will be another prospective area such as the DNA analysis in the identification of the talented children for the sports filed.

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Analysis of Interactions in Multiple Genes using IFSA(Independent Feature Subspace Analysis) (IFSA 알고리즘을 이용한 유전자 상호 관계 분석)

  • Kim, Hye-Jin;Choi, Seung-Jin;Bang, Sung-Yang
    • Journal of KIISE:Computer Systems and Theory
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    • v.33 no.3
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    • pp.157-165
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    • 2006
  • The change of external/internal factors of the cell rquires specific biological functions to maintain life. Such functions encourage particular genes to jnteract/regulate each other in multiple ways. Accordingly, we applied a linear decomposition model IFSA, which derives hidden variables, called the 'expression mode' that corresponds to the functions. To interpret gene interaction/regulation, we used a cross-correlation method given an expression mode. Linear decomposition models such as principal component analysis (PCA) and independent component analysis (ICA) were shown to be useful in analyzing high dimensional DNA microarray data, compared to clustering methods. These methods assume that gene expression is controlled by a linear combination of uncorrelated/indepdendent latent variables. However these methods have some difficulty in grouping similar patterns which are slightly time-delayed or asymmetric since only exactly matched Patterns are considered. In order to overcome this, we employ the (IFSA) method of [1] to locate phase- and shut-invariant features. Membership scoring functions play an important role to classify genes since linear decomposition models basically aim at data reduction not but at grouping data. We address a new function essential to the IFSA method. In this paper we stress that IFSA is useful in grouping functionally-related genes in the presence of time-shift and expression phase variance. Ultimately, we propose a new approach to investigate the multiple interaction information of genes.

Characterization of Bovine Lymphocyte Antigen DRB3 exon2 Gene of Korean Native Cattle (한우의 BoLA DRB3 exon2 유전자의 특성)

  • Kang, Ho Bum;Ryoo, Seung Heui;Lee, Sang Hoon;Jeon, Byung Soon;Sang, Byung Chan
    • Korean Journal of Agricultural Science
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    • v.25 no.1
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    • pp.79-88
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    • 1998
  • This study was performed in order to apply to effective breeding of Korean native cattle on the molecular genetic level obtained from PCR and nucleotide sequencing analysis of BoLA DRB3 exon2 that has important roles in host immune defence. Genomic DNA used in this study was prepared from the blood of Korean native cattle in Korean Native Cattle Improvement Center of National Livestock Cooperation. The results obtained from this study are summarized as follows: 1. Genomic DNA extracted from the blood of Korean native cattle was subjected to electrophoresis on 1.5% agarose gel. Major band was bigger than 12.2kb, indicating that genomic DNA was well prepared for PCR. Amplified products of 284bp fragments was obtained the amplification of BoLA DRB3 exon2 gene by PCR. 2. Cloning of BoLA DRB3 exon2 of Korean native cattle with pCR2.1 vector was conformed by 300bp fragment from recombinent plasmid that restricted with enzyme digestion of EcoRI. 3. Homology of BoLA DRB3 exon2 alleles of parent was 82.0% between sire's alleles and 90.1% between dam's alleles. 4. In pedigree analysis using BoLA DRB3 exon2 gene, sequencing result of BoLA DRB3 exon2 genes showed inheritance by Mendelian mode through the parents to their offspring. 5. Taking together those experimental results, pedigree was confirmed on the basis of sequencing for the alleles of parents and offspring. This knowledge by the molecular biological approach could be served for the improvement of Korean native cattle.

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