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유승민 ( Seung Min Ryu ),유승열 ( Seung Ryul Yoo ),박준석 ( Jun Seuk Park ),홍은정 ( Eun Jung Hong ),노태협 ( Tai Hyeop Lho ) 한국물환경학회 2013 한국물환경학회지 Vol.29 No.5
Chloroform is harmful volatile organics and representatives of Trihalomethane (THM). Well-known removal methods of Chloroform are photo oxidation or OH radical oxidation. Plasma on water surface at slightly vacuum condition (45 torr) can produce OH radical and it will help chloroform removal. 81.5% of chloroform is removed by vacuum and plasma in 10 min. Plasma can totally oxidize it till 2.8% and partially oxidize chloroform up to 18.5%. Water-surface plasma is good method to remove chloroform in short time.
Adenine base editing in mouse embryos and an adult mouse model of Duchenne muscular dystrophy
Ryu, Seuk-Min,Koo, Taeyoung,Kim, Kyoungmi,Lim, Kayeong,Baek, Gayoung,Kim, Sang-Tae,Kim, Heon Seok,Kim, Da-eun,Lee, Hyunji,Chung, Eugene,Kim, Jin-Soo Nature Publishing Group, a division of Macmillan P 2018 Nature biotechnology Vol.36 No.6
<P>Adenine base editors (ABEs) composed of an engineered adenine deaminase and the Streptococcus pyogenes Cas9 nickase enable adenine-to-guanine (A-to-G) single-nucleotide substitutions in a guide RNA (gRNA)-dependent manner. Here we demonstrate application of this technology in mouse embryos and adult mice. We also show that long gRNAs enable adenine editing at positions one or two bases upstream of the window that is accessible with standard single guide RNAs (sgRNAs). We introduced the Himalayan point mutation in the Tyr gene by microinjecting ABE mRNA and an extended gRNA into mouse embryos, obtaining Tyr mutant mice with an albino phenotype. Furthermore, we delivered the split ABE gene, using trans-splicing adenoassociated viral vectors, to muscle cells in a mouse model of Duchenne muscular dystrophy to correct a nonsense mutation in the Dmd gene, demonstrating the therapeutic potential of base editing in adult animals.</P>
Evolution of CRISPR towards accurate and efficient mammal genome engineering
( Seuk-min Ryu ),( Junseok W Hur ),( Kyoungmi Kim ) 생화학분자생물학회(구 한국생화학분자생물학회) 2019 BMB Reports Vol.52 No.8
The evolution of genome editing technology based on CRISPR (clustered regularly interspaced short palindromic repeats) system has led to a paradigm shift in biological research. CRISPR/Cas9-guide RNA complexes enable rapid and efficient genome editing in mammalian cells. This system induces double-stranded DNA breaks (DSBs) at target sites and most DNA breakages induce mutations as small insertions or deletions (indels) by non-homologous end joining (NHEJ) repair pathway. However, for more precise correction as knock-in or replacement of DNA base pairs, using the homology-directed repair (HDR) pathway is essential. Until now, many trials have greatly enhanced knock-in or substitution efficiency by increasing HDR efficiency, or newly developed methods such as Base Editors (BEs). However, accuracy remains unsatisfactory. In this review, we summarize studies to overcome the limitations of HDR using the CRISPR system and discuss future direction. [BMB Reports 2019; 52(8): 475-481]
Kim, Min Keun,Lee, Sun Mi,Seuk, Su Won,Ryu, Jae San,Kim, Hee Dae,Kwon, Jin Hyeuk,Choi, Yong Jo,Yun, Han Dae The Korean Society of Plant Pathology 2017 Plant Pathology Journal Vol.33 No.3
RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae. The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii.
Highly efficient RNA-guided base editing in mouse embryos
Kim, Kyoungmi,Ryu, Seuk-Min,Kim, Sang-Tae,Baek, Gayoung,Kim, Daesik,Lim, Kayeong,Chung, Eugene,Kim, Sunghyun,Kim, Jin-Soo Nature Publishing Group, a division of Macmillan P 2017 Nature biotechnology Vol.35 No.5
<P>Base editors (BEs) composed of a cytidine deaminase fused to CRISPR-Cas9 convert cytidine to uridine, leading to single-base-pair substitutions in eukaryotic cells. We delivered BE mRNA or ribonucleoproteins targeting the Dmd or Tyr gene via electroporation or microinjection into mouse zygotes. F0 mice showed nonsense mutations with an efficiency of 44-57% and allelic frequencies of up to 100%, demonstrating an efficient method to generate mice with targeted point mutations.</P>