• Journal of Plant Biotechnology
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• 제46권3호
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• pp.137-142
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• 2019

유전자교정작물은 공여 DNA의 사용 여부와 돌연변이의 크기에 따라 SDN-1, SDN-2, SDN-3 작물로 분류한다. 특히 SDN-1과 SDN-2 작물들은 이들을 창출하기 위하여 사용한 운반체 DNA 단편 또는 guide RNA가 잔존 하지 않는 100% transgene-free 작물의 개발이 가능하다. 따라서 이들은 기존의 전통교배육종기술을 이용하거나 자연 상태에서도 창출이 가능한 작물들이다. 그러므로 기존의 GMO 법령에 따라 이들 유전자교정작물을 판별하거나 표시제에 근거한 관리 감독을 수행하기가 어렵다. 이러한 과학적 사실에 근거하여 호주는 SDN-1 작물은 GMO 규제에서 제외하도록 하였다. 또한, 아르헨티나, 브라질, 일본 등은 외래유전자가 최종산물에 잔존 하지 않는 유전자교정작물은 GMO 규제에서 제외하도록 하여 SDN-1은 물론 SDN-2 작물도 GMO에 포함되지 않을 수도 있도록 하였다. 이러한 추세에 따라 우리나라도 외래유전자가 잔존 하지 않는 유전자교정작물은 GMO 규제에서 제외하도록 하여 유전자교정기술을 이용한 다양한 작물 육종 계통 육성이 널리 이용되어 우수 품종 육성에 기여되길 기대한다.

• Experimental and Molecular Medicine
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• 제50권12호
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• pp.1.1-1.14
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• 2018

DNA methylation is a regulatory mechanism in epigenetics that is frequently altered during human carcinogenesis. To detect critical methylation events associated with gastric cancer (GC), we compared three DNA methylomes from gastric mucosa (GM), intestinal metaplasia (IM), and gastric tumor (GT) cells that were microscopically dissected from an intestinal-type early gastric cancer (EGC) using methylated DNA binding domain sequencing (MBD-seq) and reduced representation bisulfite sequencing (RRBS) analysis. In this study, we focused on differentially methylated promoters (DMPs) that could be directly associated with gene expression. We detected 2,761 and 677 DMPs between the GT and GM by MBD-seq and RRBS, respectively, and for a total of 3,035 DMPs. Then, 514 (17%) of all DMPs were detected in the IM genome, which is a precancer of GC, supporting that some DMPs might represent an early event in gastric carcinogenesis. A pathway analysis of all DMPs demonstrated that 59 G protein-coupled receptor (GPCR) genes linked to the hypermethylated DMPs were significantly enriched in a neuroactive ligand-receptor interaction pathway. Furthermore, among the 59 GPCRs, six GI hormone receptor genes (NPY1R, PPYR1, PTGDR, PTGER2, PTGER3, and SSTR2) that play an inhibitory role in the secretion of gastrin or gastric acid were selected and validated as potential biomarkers for the diagnosis or prognosis of GC patients in two cohorts. These data suggest that the loss of function of gastrointestinal (GI) hormone receptors by promoter methylation may lead to gastric carcinogenesis because gastrin and gastric acid have been known to play a role in cell differentiation and carcinogenesis in the GI tract.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.13-13
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• 2017

By the progress of genome sequencing, infrastructures for marker-assisted breeding (MAB) of rice came to be established. Fine mapping and gene isolation have been conducted using the breeding materials derived from natural variations and artificial mutants. Such genetic analysis by the genome-wide dense markers provided us the knowledge about the many genes controlling important traits. We identified several genes or quantitative trait loci (QTL) for heading date, blast resistance, eating quality, high-temperature stress tolerance, and so on. NILs of each gene controlling heading date contribute to elongate the rice harvest period. Determination of precise gene location of blast resistance gene pi21, allowed us to overcome linkage drag, co-introduction of undesirable eating quality. We could also breed the first practical rice cultivar in Japan with a brown planthopper resistance gene bph11 in the genetic back-ground of an elite cultivar. Discovery of major and minor QTLs for good eating quality allowed us to fine-tune of eating quality according to the rice planting area or usage of rice grain. Many rice cultivars have bred efficiently by MAB for several traits, or by marker-assisted backcross breeding through chromosome segment substitution lines (CSSLs) using genetically diverse accessions. We are also systematically supporting the crop breeding of other sectors by MAB or by providing resources such as CSSLs. It is possible to pyramid many genes for important traits by using MAB, but is still difficult to improve the yielding ability. We are performing a Genomic Selection (GS) for improvement of rice biomass and grain yield. We are also trying to apply the genome editing technology for high yield rice breeding.

• Journal of Genetic Medicine
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• 제20권2호
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• pp.39-45
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• 2023

As advanced sequencing technologies continue to uncover an increasing number of variants in genes associated with human genetic diseases, there is a growing demand for systematic approaches to assess the impact of these variants on human development, health, and disease. While in silico analyses have provided valuable insights, it is essential to complement these findings with model organism studies to determine the functional consequences of genetic variants in vivo. Drosophila melanogaster is an excellent genetic model for such functional studies due to its efficient genetic technologies, high gene conservation with humans, accessibility to mutant fly resources, short life cycles, and cost-effectiveness. The traditional GAL4-UAS system, allowing precise control of gene expression through binary regulation, is frequently employed to assess the effects of monoallelic variants. Recombinase medicated cassette exchange or CRISPR-Cas9-mediated GAL4 insertion within coding introns or substitution of gene body with Kozak-Gal4 result in the loss-of-function of the target gene. This GAL4 insertion strategy also enables the expression of reference complementary DNA (cDNA) or cDNA carrying genetic variants under the control of endogenous regulatory cis elements. Furthermore, the CRISPR-Cas9-directed tissue-specific knockout and cDNA rescue system provides the flexibility to investigate candidate variants in a tissue-specific and/or developmental-timing dependent manner. In this review, we will delve into the diverse genetic techniques available in Drosophila and their applications in diagnosing and studying numerous undiagnosed diseases over the past decade.

• 생명과학회지
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• 제30권8호
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• pp.701-707
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• 2020

Clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR associated protein (Cas)9을 이용하는 유전자 가위 기술은 미래 육종 기술로서 주목받고 있다. 본 연구에서는 3세대 유전자 가위 CRISPR/Cas9을 이용한 누에 kynurenine 3-monooxygenase (KMO) 유전자 편집을 통한 돌연변이 유발 및 선택 교배를 통하여 유전자의 세대간 전달을 분석하고자 하였다. 유전자 편집을 위하여 누에의 KMO 유전자에 대한 3종의 가이드 RNA를 제작하고, 제작된 gRNA는 Cas9 단백질과 복합체를 형성시켜 누에 세포주(BM-N)에 도입 후 T7 endonuclease I 분석을 수행하여 최적의 gRNA를 선발하였다. 선발된 K1N gRNA는 누에 유전자를 편집하기 위하여 Cas9 단백질과 복합체를 형성시킨 후 누에 초가 배아에 미세주사하고 사육하였다. 미세주사 후 부화율은 18% 가량으로 낮게 나타났으나 생존한 개체 중 돌연변이 발생율은 60% 이상으로 비교적 높게 나타났다. 돌연변이가 발생된 G0세대의 KMO 유전자는 이형접합자 형태로 나타났으며, 표현형의 변화는 관찰되지 않았다. 하지만 이형접합자들 사이의 근친 교배에 의해 탄생한 G1세대 돌연변이에서는 일부에서 알과 눈의 색 변화가 확인되었으며, 변이가 확인된 개체들사이의 근친교배를 통해 생산된 G2세대에서는 모든 개체에서 표현형의 변화가 나타났다. 이러한 결과에 비추어 볼 때, 유전자 가위를 이용한 돌연변이 육종에는 한계가 있으나 전통 교배 육종과 융합을 통하여 육종 기간을 비약적으로 단축시킬 수 있는 곤충 육종 기술로 발전가능성이 높다고 판단된다.

• Genomics & Informatics
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• 제18권4호
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• pp.46.1-46.4
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• 2020

We developed the BaSDAS (Barcode-Seq Data Analysis System), a GUI-based pooled knockout screening data analysis system, to facilitate the analysis of pooled knockout screen data easily and effectively by researchers with limited bioinformatics skills. The BaSDAS supports the analysis of various pooled screening libraries, including yeast, human, and mouse libraries, and provides many useful statistical and visualization functions with a user-friendly web interface for convenience. We expect that BaSDAS will be a useful tool for the analysis of genome-wide screening data and will support the development of novel drugs based on functional genomics information.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2022년도 추계학술대회
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• pp.285-285
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• 2022

Recent unpredictable climate change is a major cause of rice yield loss. In particular, methane is a key factor in global warming. Therefore rice breeders are trying to breed the reducing-methane gas emission rice using the crossbreeding method. However, the traditional crossbreeding method takes 8 to 10 years to breed a cultivar, and the anther culture method developed to shorten the breeding cycle also takes 6 to 7 years. On the other hand, CRISPR/Cas9 accurately edits the target trait and can rapidly breed rice cultivars by editing the target trait as a homozygous in 2-3 years. In addition, exogenous genetic elements such as Cas9 can be isolated from the G₁ generation. Therefore, the flowering time was regulated by applying CRISPR/Cas9 technology, and OsCKq1 genome-editing (OsCKq1-G) rice with early flowered and high yield was bred in the field. Genome-editing of OsCKq1 applied CRISPR/Cas9 technology up-regulates the expression of the flowering promotion gene Ehd1 under long-day conditions induces early flowering and increases the yield by increasing the 1,000-grain weight. And as the generations advanced, each agricultural trait indicated a low coefficient of variation. As a result, indicated that OsCKq1 plays an important role in regulating the flowering time and is related to the trait determining yield. Therefore, OsCKq1-G can suggest a breeding strategy for the Net-Zero national policy for reducing-methane gas emission rice by shortening the breeding cycle with the early flowered, and high-yield rice. CRISPR/Cas9 technology is a rapid and accurate breeding technology for breeding rice cultivars with important characteristics.

• Molecules and Cells
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• 제38권9호
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• pp.743-749
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• 2015

Production of genome-edited animals using germline-competent cells and genetic modification tools has provided opportunities for investigation of biological mechanisms in various organisms. The recently reported programmed genome editing technology that can induce gene modification at a target locus in an efficient and precise manner facilitates establishment of animal models. In this regard, the demand for genome-edited avian species, which are some of the most suitable model animals due to their unique embryonic development, has also increased. Furthermore, germline chimera production through longterm culture of chicken primordial germ cells (PGCs) has facilitated research on production of genome-edited chickens. Thus, use of avian germline modification is promising for development of novel avian models for research of disease control and various biological mechanisms. Here, we discuss recent progress in genome modification technology in avian species and its applications and future strategies.

• 식물분류학회지
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• 제52권2호
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• pp.114-117
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• 2022

Lindera angustifolia is mainly distributed in the temperate climate zone of China but shows an extraordinary distribution, disjunctively isolated on the western coastal islands of Korea. We therefore present the complete chloroplast genome of Korean L. angustifolia. The complete plastome was 152,836 bp in length, with an overall GC content of 39.2%. A large single copy (93,726 bp) and a small single copy (18,946 bp) of the genome were separated by a pair of inverted repeats (20,082 bp). The genome consists of 125 genes, including 81 protein-coding, eight ribosomal RNA, and 36 transfer RNA genes. While five RNA editing genes (psbL, rpl2, ndhB×2, and ndhD) were identified in L. angustifolia from China, the "ndhD" gene was not recognized as an RNA editing site in the corresponding Korean individual. A phylogenetic analysis revealed that Korean L. angustifolia is most closely related to the Chinese L. angustifolia with strong bootstrap support, forming a sister group of L. glauca.

• 한국미생물·생명공학회지
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• 제51권4호
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• pp.374-389
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• 2023

Organophosphorus (OP) pesticides, particularly malathion, parathion, diazinon, and chlorpyrifos, are widely used in both agricultural and residential contexts. This refractory quality is shared by certain organ phosphorus insecticides, and it may have unintended consequences for certain non-target soil species. Bioremediation cleans organic and inorganic contaminants using microbes and plants. Organophosphate-hydrolyzing enzymes can transform pesticide residues into non-hazardous byproducts and are increasingly being considered viable solutions to the problem of decontamination. When coupled with system analysis, the multi-omics technique produces important data for functional validation and genetic manipulation, both of which may be used to boost the efficiency of bioremediation systems. RNA-guided nucleases and RNA-guided base editors include zinc-finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and clustered regularly interspaced short palindromic repeats (CRISPR), which are used to alter genes and edit genomes. The review sheds light on key knowledge gaps and suggests approaches to pesticide cleanup using a variety of microbe-assisted methods. Researches, ecologists, and decision-makers can all benefit from having a better understanding of the usefulness and application of systems biology and gene editing in bioremediation evaluations.