• 제목/요약/키워드: Prime editing

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Genome-wide in-locus epitope tagging of Arabidopsis proteins using prime editors

  • Cheljong Hong;Jun Hee Han;Gue-Ho Hwang;Sangsu Bae;Pil Joon Seo
    • BMB Reports
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    • 제57권1호
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    • pp.66-70
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    • 2024
  • Prime editors (PEs), which are CRISPR-Cas9 nickase (H840A)-reverse transcriptase fusion proteins programmed with prime editing guide RNAs (pegRNAs), can not only edit bases but also install transversions, insertions, or deletions without both donor DNA and double-strand breaks at the target DNA. As the demand for in-locus tagging is increasing, to reflect gene expression dynamics influenced by endogenous genomic contexts, we demonstrated that PEs can be used to introduce the hemagglutinin (HA) epitope tag to a target gene locus, enabling molecular and biochemical studies using in-locus tagged plants. To promote genome-wide in-locus tagging, we also implemented a publicly available database that designs pegRNAs for in-locus tagging of all the Arabidopsis genes.

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

  • Min-cheol Kim;Tae-Hwan Jun
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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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
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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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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RNA in Therapeutics: CRISPR in the Clinic

  • Dana Carroll
    • Molecules and Cells
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    • 제46권1호
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    • pp.4-9
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    • 2023
  • The advent of the CRISPR-Cas genome editing platform has greatly enhanced the capabilities of researchers in many areas of biology. Its use has also been turned to the development of therapies for genetic diseases and to the enhancement of cell therapies. This review describes some recent advances in these areas.

Applications of CRISPR technologies to the development of gene and cell therapy

  • Chul-Sung Park;Omer Habib;Younsu Lee;Junho K. Hur
    • BMB Reports
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    • 제57권1호
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    • pp.2-11
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    • 2024
  • Advancements in gene and cell therapy have resulted in novel therapeutics for diseases previously considered incurable or challenging to treat. Among the various contributing technologies, genome editing stands out as one of the most crucial for the progress in gene and cell therapy. The discovery of CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) and the subsequent evolution of genetic engineering technology have markedly expanded the field of target-specific gene editing. Originally studied in the immune systems of bacteria and archaea, the CRISPR system has demonstrated wide applicability to effective genome editing of various biological systems including human cells. The development of CRISPR-based base editing has enabled directional cytosine-to-thymine and adenine-to-guanine substitutions of select DNA bases at the target locus. Subsequent advances in prime editing further elevated the flexibility of the edit multiple consecutive bases to desired sequences. The recent CRISPR technologies also have been actively utilized for the development of in vivo and ex vivo gene and cell therapies. We anticipate that the medical applications of CRISPR will rapidly progress to provide unprecedented possibilities to develop novel therapeutics towards various diseases.

Therapeutic applications of gene editing in chronic liver diseases: an update

  • Shin, Ji Hyun;Lee, Jinho;Jung, Yun Kyung;Kim, Kyeong Sik;Jeong, Jaemin;Choi, Dongho
    • BMB Reports
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    • 제55권6호
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    • pp.251-258
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    • 2022
  • Innovative genome editing techniques developed in recent decades have revolutionized the biomedical research field. Liver is the most favored target organ for genome editing owing to its ability to regenerate. The regenerative capacity of the liver enables ex vivo gene editing in which the mutated gene in hepatocytes isolated from the animal model of genetic disease is repaired. The edited hepatocytes are injected back into the animal to mitigate the disease. Furthermore, the liver is considered as the easiest target organ for gene editing as it absorbs almost all foreign molecules. The mRNA vaccines, which have been developed to manage the COVID-19 pandemic, have provided a novel gene editing strategy using Cas mRNA. A single injection of gene editing components with Cas mRNA is reported to be efficient in the treatment of patients with genetic liver diseases. In this review, we first discuss previously reported gene editing tools and cases managed using them, as well as liver diseases caused by genetic mutations. Next, we summarize the recent successes of ex vivo and in vivo gene editing approaches in ameliorating liver diseases in animals and humans.

인터렉티브 라이브모션 저작도구 Motion Space Prime (Interactive Live Motion Authoring Tool : Motion Space Prime)

  • 박승규;이황기;진성아
    • 한국콘텐츠학회:학술대회논문집
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    • 한국콘텐츠학회 2003년도 춘계종합학술대회논문집
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    • pp.107-110
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    • 2003
  • 최근 애니메이션이 대중에게 널리 보급되면서 애니메이션을 제작 편집 하는 분야가 크게 각광 받고 있다. 그럼에도 불구하고 리눅스 환경에서 애니메이션분야의 편집은 윈도우 환경에 비해 미비한 실정이다. 본 논문에서는 보다 편하고 쉽게 Clay animation과 같은 stop motion animation을 리눅스 환경에서도 원활하게 제작 할 수 있는 Motion Space Prime을 제안한다. Motion Space Prime을 이용하여 디지털 캠코더의 영상을 실시간으로 캡쳐하고 사용자가 인터렉티브하게 편집하여 애니메이션을 쉽게 제작 할 수 있다.

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Development of CRISPR technology for precise single-base genome editing: a brief review

  • Lee, Hyomin K.;Oh, Yeounsun;Hong, Juyoung;Lee, Seung Hwan;Hur, Junho K.
    • BMB Reports
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    • 제54권2호
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    • pp.98-105
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    • 2021
  • The clustered regularly interspaced short palindromic repeats (CRISPR) system is a family of DNA sequences originally discovered as a type of acquired immunity in prokaryotes such as bacteria and archaea. In many CRISPR systems, the functional ribonucleoproteins (RNPs) are composed of CRISPR protein and guide RNAs. They selectively bind and cleave specific target DNAs or RNAs, based on sequences complementary to the guide RNA. The specific targeted cleavage of the nucleic acids by CRISPR has been broadly utilized in genome editing methods. In the process of genome editing of eukaryotic cells, CRISPR-mediated DNA double-strand breaks (DSB) at specific genomic loci activate the endogenous DNA repair systems and induce mutations at the target sites with high efficiencies. Two of the major endogenous DNA repair machineries are non-homologous end joining (NHEJ) and homology-directed repair (HDR). In case of DSB, the two repair pathways operate in competition, resulting in several possible outcomes including deletions, insertions, and substitutions. Due to the inherent stochasticity of DSB-based genome editing methods, it was difficult to achieve defined single-base changes without unanticipated random mutation patterns. In order to overcome the heterogeneity in DSB-mediated genome editing, novel methods have been developed to incorporate precise single-base level changes without inducing DSB. The approaches utilized catalytically compromised CRISPR in conjunction with base-modifying enzymes and DNA polymerases, to accomplish highly efficient and precise genome editing of single and multiple bases. In this review, we introduce some of the advances in single-base level CRISPR genome editing methods and their applications.

'융·복합'과 디드로의 철학 - 『자연해석론』을 중심으로 - (Interdisciplinary and philosophy of Diderot Considered through De l'interprétation de la nature)

  • 백찬욱
    • 비교문화연구
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    • 제33권
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    • pp.143-169
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    • 2013
  • Recently, because of the danger and damages that following the steady development of scientific technologies, natural science is faced with many humane and ethical problems. So it is asserted the interdisciplinary with social and human science. However, it comes commonly that the ethical issues such as world view and one's view of life caused the development of sociology, especially development of the market economy. But in many cases, the interdisciplinary can be useful for widen the view of scholars. The interdisciplinary is actually connected with the problems of philosophy, and located in that domain. In this case, this is noticed as a model to the philosophers in 18th century, especially Denis Diderot. Diderot published De $l^{\prime}interpr{\acute{e}}tation$ de la nature during editing the Encyclopedia, from there he picked out the contents from piles of documents of Encyclopedia. Even though the contents or opinions of De $l^{\prime}interpr{\acute{e}}tation$ de la nature are inaccuracy or erroneous, it shows that how human-social science and natural science encountered. Diderot studied mathematics and then Diderot accepted to the natural science proposal, he approaches philosophy with translate English books to the French. Next he understood natural science by reading Buffon and Maupertuis, and during working for Encyclopedia, he possessed his knowledge that he can claim his opinion to other scholars. However in this De $l^{\prime}interpr{\acute{e}}tation$ de la nature, Diderot who sometimes rebutted other scholars' theory and demonstration, tried to build a philosophy on metaphysics in order to it was important for himself that he imposed the methods of science and importance of experience. Anyhow, this De $l^{\prime}interpr{\acute{e}}tation$ de la nature cause consider the recognition of Diderot in the field of natural science, and is suggested as a model about his Nature. This mean that it is an expression of his philosophy, and the content is found from natural philosophy and empirical philosophy. Like giving these attache the importance of method study for science and technique, these are targeted the promotion of popularization of natural-science and scientific-technology. Also it advocates fulfilling from reasonable philosophy to empirical philosophy. Therefore, the philosophy which was speculative and abstracted became his philosophy which was writing the meaning, as waiting the discovery of science. And at that time, the humanities made interdisciplinary with natural science.

사부송유목록에 대한 연구 (A study on the Sabu-Songyu-Mongnok(사부송유목록), or a Classified Catalogue for Recitation and Appreciation of Classics)

  • 이상용
    • 한국문헌정보학회지
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    • 제25권
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    • pp.445-476
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    • 1993
  • This paper is written to clarify the specific details of the Sabu-Songyu­Mongnok(사부송유목록). which is a reading list for recitation and appreciation of Chinese classics edited by Hong Suk-Chu(1774-1842) when he was 56 years old for the stimulation of his younger brother Hyun-Ju (1793-1865)'s reading life. In this study the catalogue's title interpretation. the time of the editing, the motive for the editing and the analysis of the contents are revealed as what they actually are. The main details are as followings. There is no date written on the Sabu-Songyu-Mongnok but the writer estimate it to be around 1829. After Hong Suk-Chu recognized the fact that although his younger brother Hyun-Ju was interested in reading but felt that he was a little too old to begin to read all the books of the whole classes. he selected the classic works from whole subjects, i.e. history, philosophy and anthology, that he thought which would best represent each subject. From these selected works he then picked out the most important chapters or parts of the works. Another motivation of the editing was for the stimulation of the reading life for Hong Suk-chu himself and for people like himself who were getting old and losing their vitality. The Sabu-Songyu-Mongnok consists of four categories. i.e. classics category, history category, philosophy category and anthology category, and in the catalogue, 26 different works are entered. The classics category has ten, the history category has six, the philosophy category has six and the anthology category has four. Each of the works are entered under the title of the book, and hen the chapter heading. The Sabu-Songyu-Mongnok was edited 19 years later than Hong-ssi toksorok(홍씨독서록) or an Annotated Bibliography of Korean and Chinese Books. Comparing each category of the catalogue with the Hong-ssi toksorok, the writer could catch the following characteristics. 1. Although there was no indication of the class heading, the sequence of the arrangement of the entries in the classics categories was the same as the sequence of the classes in the Hong-ssi toksorok. 2. The Zuo-shi Chun-qiu-zhuan(좌씨춘추전) is recorded in the class of Chunqiu of the classics category in the Hong-ssi toksorok, but in the Sabu-Songyu­Mongnok, it belongs to the history category. 3. In the philosophy category of the catalogue, Chuangtzu(장자) writings are included because it is so well written. 4. Unlike the categories of classics, history and anthology, the philosophy category is recorded under the author's name. This is fascinating because it unites with the conventional cataloguing practice of the west. 5. In the anthology category, the writings with the best styles from the Chuci(초사) or the poems of Chou, Wen-xuan(문선), wen-yuan-ying-hua(문여영화) are selected and classified into four parts: (1) poems, (2) proses of Han, Wei and Pre-Qin dynasties. (3) Si-liu-pian-li-wen(사육병려문), (4) classic style of writings of Tang and Song dynasties (당송고문). 6. There was an unusually large number of Han Yu's writings selected from the great eight poets of Tang and Song dynasty writings. (20 categories out of $43: 47\%)$ After comparing the entries of the Sabu-Songyu-Mongnok, it can be concluded that all the entries were also included in the Hong-ssi tokrorok. The fact that the sequence of the entries in the classics category and that the literary works were sequenced under the same subject by the year of editing or writing like as the Hong-ssi toksorok Also it is unusual that there was not even one literary work of a Korean in the catalogue. Anyhow this catalogue is the first recitation catalogue in our country that was edited for the people in the prime of life as well as for old people and is the only one existing today.

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