• Title/Summary/Keyword: editing

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Gene Editing for Major Allergy Genes using Multiplex CRISPR-Cas9 System & Prime editing in Peanuts (Arachis hypogaea L.)

  • Min-cheol Kim;Tae-Hwan Jun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.194-194
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    • 2022
  • Recently, food-induced allergies have emerged as major global concerns. In the past ten years, it has doubled in western nations, and it has also increased in Asia and Africa. In many cases of food allergy, peanut allergy is prevalent, typically permanent, and frequently life-threatening. Therefore, we utilized gene editing techniques on the three major allergen genes in peanuts, Ara h 1, Ara h 2, and Ara h 3. Using gibson assembly and golden gate assembly, we created two vectors, the gRNA-tRNA array CRISPR-Cas9 system and Prime-editing. Using LBA4404 strain and agrobacterium-mediated transformation, the vectors were transferred to two elite Korean peanut lines. After co-cultivation and tissue culture, we extracted the tissue cultured peanut DNA amplified the hygromycin resistance gene and Cas9 gene in the T-DNA region. The integration of the T-DNA region into the host genome was demonstrated by the presence of a specific band in some samples. There have only been a few reported peanut gene editing studies. So, this study will contribute to peanut allergy and gene editing research.

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Gene Editing for Major Allergy Genes using Multiplex CRISPR-Cas9 System & Prime Editing in Peanuts (Arachis hypogaea L.)

  • Min-cheol Kim;Tae-Hwan Jun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.200-200
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    • 2022
  • Recently, food-induced allergies have emerged as major global concerns. In the past ten years, it has doubled in western nations, and it has also increased in Asia and Africa. In many cases of food allergy, peanut allergy is prevalent, typically permanent, and frequently life-threatening. Therefore, we utilized gene editing techniques on the three major allergen genes in peanuts, Ara h 1, Ara h 2, and Ara h 3. Using gibson assembly and golden gate assembly, we created two vectors, the gRNA-tRNA array CRISPR-Cas9 system and Prime-editing. Using LBA4404 strain and agrobacterium-mediated transformation, the vectors were transferred to two elite Korean peanut lines. After co-cultivation and tissue culture, we extracted the tissue cultured peanut DNA amplified the hygromycin resistance gene and Cas9 gene in the T-DNA region. The integration of the T-DNA region into the host genome was demonstrated by the presence of a specific band in some samples. There have only been a few reported peanut gene editing studies. So, this study will contribute to peanut allergy and gene editing research.

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Mitochondrial genome editing: strategies, challenges, and applications

  • Kayeong Lim
    • BMB Reports
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    • v.57 no.1
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    • pp.19-29
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    • 2024
  • Mitochondrial DNA (mtDNA), a multicopy genome found in mitochondria, is crucial for oxidative phosphorylation. Mutations in mtDNA can lead to severe mitochondrial dysfunction in tissues and organs with high energy demand. MtDNA mutations are closely associated with mitochondrial and age-related disease. To better understand the functional role of mtDNA and work toward developing therapeutics, it is essential to advance technology that is capable of manipulating the mitochondrial genome. This review discusses ongoing efforts in mitochondrial genome editing with mtDNA nucleases and base editors, including the tools, delivery strategies, and applications. Future advances in mitochondrial genome editing to address challenges regarding their efficiency and specificity can achieve the promise of therapeutic genome editing.

The Design and Implementation of SGML Document Editing System Using Document Structure Information (문서 구조정보를 이용한 SGML 문서 편집 시스템의 설계 및 구현)

  • Kim, Chang-Su;Jo, In-June;Jung, Hoe-Kyung
    • The Journal of Engineering Research
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    • v.3 no.1
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    • pp.21-27
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    • 1998
  • This paper describes the design and implementation of system for editing SGML document instance using document structure information of SGML DTD. For make use of structure window for logical structure expression of document to SGML document editing without editing mistake of user and easy update the using support to editing process of elements, attributes, entities tools and product document, and valid using SGML parser. Also, in order to support Korean and English text using KS 5601. In this paper, the proposed SGML document editing system is used common controls support of window 95 for window user interface

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Editing of Genomic TNFSF9 by CRISPR-Cas9 Can Be Followed by Re-Editing of Its Transcript

  • Lee, Hyeon-Woo
    • Molecules and Cells
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    • v.41 no.10
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    • pp.917-922
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    • 2018
  • The CRISPR-Cas system is a well-established RNA-guided DNA editing technique widely used to modify genomic DNA sequences. I used the CRISPR-Cas9 system to change the second and third nucleotides of the triplet $T{\underline{CT}}$ of human TNSFSF9 in HepG2 cells to $T{\underline{AG}}$ to create an amber stop codon. The $T{\underline{CT}}$ triplet is the codon for Ser at the $172^{nd}$ position of TNSFSF9. The two substituted nucleotides, AG, were confirmed by DNA sequencing of the PCR product followed by PCR amplification of the genomic TNFSF9 gene. Interestingly, sequencing of the cDNA of transcripts of the edited TNFSF9 gene revealed that the $T{\underline{AG}}$ had been re-edited to the wild type triplet $T{\underline{CT}}$, and 1 or 2 bases just before the triplet had been deleted. These observations indicate that CRISPR-Cas9-mediated editing of bases in target genomic DNA can be followed by spontaneous re-editing (correcting) of the bases during transcription.

Molecular cDNA cloning and unusual RNA editings of NAD4 gene from Zea mays mitochondrion (옥수수 미토콘드리아 NAD4유전자의 cDNA cloning과 특이한 RNA editing 현상)

  • 설일환
    • Journal of Life Science
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    • v.8 no.2
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    • pp.203-207
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    • 1998
  • NAD4 as a gene encoding NADH dehydrogenase subunit 4 in the micodhondrion from maize has been cloned using RT-PCR and sequenced for examining RNA edited sites. Analysis of mt cDNA sequences showed the typical RNA editing patterns and unusual base changes as well;RNA editing from cDNA sequences occured base change from c to U in most cases, however transitions from t to g and G to A were also observed. Even though those editings appared to be occurred randomly, RNA edited sites showed mostly in exon 1 and exon 4 regions, when compared with NAD4 cDNA from wheat, locations of edited sites did not consistent with each other suggesting that the phenomenon of RNA editing occured randomly not site-specific manner.

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Three Common Subunits in Editing Domains of Class Ia tRNA Synthetases

  • Lee, Keun-Woo;Kwon, Yong-Jung;Briggs, James M.
    • Bulletin of the Korean Chemical Society
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    • v.28 no.2
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    • pp.207-210
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    • 2007
  • To identify structural or functional common subunit(s) in the CP1 (editing) domains of class Ia tRNA synthetases, five available structures were compared and analyzed. Through the sequence alignments and structural overlapping of the CP1 domains, three conserved regions were identified near the amino acid binding site in the editing domain. Structural overlapping of the three subunits clearly showed the existence of three common structural subunits in all of the five editing RS structures. Based on the established experimental results and our modeling results, it is proposed that subunits 1 and 3 accommodate the incoming amino acid binding, while subunit 2 contributes to the interactions with the adenosine ring of the A76 to stabilize the overall tRNA binding. Since these subunits are critical for the editing reaction, we expect that these key structures should be conserved through the most class Ia editing RSs.

RPS5A Promoter-Driven Cas9 Produces Heritable Virus-Induced Genome Editing in Nicotiana attenuata

  • Oh, Youngbin;Kim, Sang-Gyu
    • Molecules and Cells
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    • v.44 no.12
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    • pp.911-919
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    • 2021
  • The virus-induced genome editing (VIGE) system aims to induce targeted mutations in seeds without requiring any tissue culture. Here, we show that tobacco rattle virus (TRV) harboring guide RNA (gRNA) edits germ cells in a wild tobacco, Nicotiana attenuata, that expresses Streptococcus pyogenes Cas9 (SpCas9). We first generated N. attenuata transgenic plants expressing SpCas9 under the control of 35S promoter and infected rosette leaves with TRV carrying gRNA. Gene-edited seeds were not found in the progeny of the infected N. attenuata. Next, the N. attenuata ribosomal protein S5 A (RPS5A) promoter fused to SpCas9 was employed to induce the heritable gene editing with TRV. The RPS5A promoter-driven SpCas9 successfully produced monoallelic mutations at three target genes in N. attenuata seeds with TRV-delivered guide RNA. These monoallelic mutations were found in 2%-6% seeds among M1 progenies. This editing method provides an alternative way to increase the heritable editing efficacy of VIGE.

Recent advances in genome engineering by CRISPR technology

  • Youngsik Lee;Yeounsun Oh;Seung Hwan Lee
    • BMB Reports
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    • v.57 no.1
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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.

A Case Study of Data Editing for the Korean Housing Price Survey (주택가격동향조사를 위한 데이터편집 사례연구)

  • Park, Jin-Woo;Park, Hyun-Joo;Kim, Jin-Eok
    • Survey Research
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    • v.6 no.1
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    • pp.83-98
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    • 2005
  • Large scale survey database may contain some erroneous data or missing data. Incomplete or erroneous data may be produced in the process of data collection or data capture. Since erroneous data can cause some bias and inconsistency, data editing, which is the procedure for detecting and adjusting individual errors in data records, is a very important work in statistical survey. In this paper, we introduce an editing process for the housing price survey to enhance discussions on that topic. We explain how to decide some appropriate edit rules and show some related data. Furthermore, we describe input editing procedures which is appropriate for on-line survey and how to find and eliminate erroneous data through output editing.

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