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Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
Moon, Suyun,An, Jee Young,Choi, Yeon-Jae,Oh, Youn-Lee,Ro, Hyeon-Su,Ryu, Hojin The Korean Society of Mycology 2021 韓國菌學會誌 Vol.49 No.6
CRISPR/Cas9 genome editing systems have been established in a broad range of eukaryotic species. Herein, we report the first method for genetic engineering in pyogo (shiitake) mushrooms (Lentinula edodes) using CRISPR/Cas9. For in vivo expression of guide RNAs (gRNAs) targeting the mating-type gene HD1 (LeA1), we identified an endogenous LeU6 promoter in the L. edodes genome. We constructed a plasmid containing the LeU6 and glyceraldehyde-3-phosphate dehydrogenase (LeGPD) promoters to express the Cas9 protein. Among the eight gRNAs we tested, three successfully disrupted the LeA1 locus. Although the CRISPR-Cas9-induced alleles did not affect mating with compatible monokaryotic strains, disruption of the transcription levels of the downstream genes of LeHD1 and LeHD2 was detected. Based on this result, we present the first report of a simple and powerful genetic manipulation tool using the CRISPR/Cas9 toolbox for the scientifically and industrially important edible mushroom, L. edodes.
Construction of a CRISPR/Cas9-Mediated Genome Editing System in Lentinula edodes
( Suyun Moon ),( Jee Young An ),( Yeon-jae Choi ),( Youn-lee Oh ),( Hyeon-su Ro ),( Hojin Ryu ) 한국균학회 2021 Mycobiology Vol.49 No.6
CRISPR/Cas9 genome editing systems have been established in a broad range of eukaryotic species. Herein, we report the first method for genetic engineering in pyogo (shiitake) mushrooms (Lentinula edodes) using CRISPR/Cas9. For in vivo expression of guide RNAs (gRNAs) targeting the mating-type gene HD1 (LeA1), we identified an endogenous LeU6 promoter in the L. edodes genome. We constructed a plasmid containing the LeU6 and glyceraldehyde-3-phosphate dehydrogenase (LeGPD) promoters to express the Cas9 protein. Among the eight gRNAs we tested, three successfully disrupted the LeA1 locus. Although the CRISPRCas9-induced alleles did not affect mating with compatible monokaryotic strains, disruption of the transcription levels of the downstream genes of LeHD1 and LeHD2 was detected. Based on this result, we present the first report of a simple and powerful genetic manipulation tool using the CRISPR/Cas9 toolbox for the scientifically and industrially important edible mushroom, L. edodes.
Suyun Moon,Hojin Ryu 한국균학회 2022 韓國菌學會誌 Vol.50 No.3
Pyogo (Shiitake, Lentinula edodes) is one of the most important edible mushrooms because of its outstanding nutritive and medicinal value. In the registration and protection procedure for newly developed mushroom cultivars, the application of molecular markers that can supplement the morphological characteristic-based distinction has been strongly requested. Sanjo 701 and Chamaram, newly developed at the Federation Forest Mushroom Research Center of Korea, have been characterized as innovative cultivars suitable for customer demands because of their high yields and cultivation rates. However, no technical tools can protect the rights to these important cultivars. In this study, using comparative genomic information from 23 commercially available pyogo cultivars, we identified single nucleotide polymorphisms (SNPs) that accurately differentiated Sanjo701 and Chamaram from the other cultivars. We also developed high-resolution melting analysis (HRM)-based SNP markers that discriminate among the tested 23 pyogo cultivars. The developed SNP markers can be utilized for rapid, accurate identification of pyogo cultivars with low genetic diversity and to prevent cultivar contamination caused by illegally distributed inocula. In addition, these markers can serve as a crucial scientific basis for securing the right to conserve new cultivars in international markets.
적응형 소프트웨어 개발을 위한 문맥 기반 요구사항 분석 방법
장호진(Hojin Jang),문미경(Mikyeong Moon),염근혁(Keunhyuk Yeom) 한국정보과학회 2004 한국정보과학회 학술발표논문집 Vol.31 No.2Ⅱ
소프트웨어의 외부 환경이 동적으로 변화하고 복잡해지면서 소프트웨어가 예상하지 못한 외부 환경의 변화 에 직면하였을 때 변화를 감지하고 대안을 선택하여 지속적인 서비스를 제공할 필요성이 증가하고 있다. 이를 위해 외부 환경의 변화를 감지하고 변화에 적응할 수 있는 적응형 소프트웨어가 나오게 되었다. 그러나 적응형 소프트웨어를 개방하고자 할 때 기존의 요구사항 분석 방법은 소프트웨어의 외부 환경의 변화에 대한 고려가 부족하다. 본 논문에서는 적응형 소프트웨어의 외부 환경의 변화와 그러한 변화에 의해 가변적으로 나타나는 요구사항을 분석하기 위한 문맥 기반 요구사항 분석 방법을 제시한다.
Lee, HoJin,Hong, Jae Taek,Kim, Il Sup,Kim, Moon Suk,Sung, Jae Hoon,Lee, Sang Won The Korean Neurosurgical Society 2014 Journal of Korean neurosurgical society Vol.56 No.6
Objective : To evaluate the anatomical feasibility of 3.5 mm screw into the cervical spine in the pediatric population and to establish useful guidelines for their placement. Methods : A total of 37 cervical spine computerized tomography scans (24 boys and 13 girls) were included in this study. All patients were younger than 10 years of age at the time of evaluation for the period of 2007-2011. Results : For the C1 screw placement, entry point height (EPH) was the most restrictive factor (47.3% patients were larger than 3.5 mm). All C2 lamina had a height larger than 3.5 mm and 68.8% (51/74) of C2 lamina had a width thicker than 3.5 mm. For C2 pedicle width, 55.4% (41/74) of cases were larger than 3.5 mm, while 58.1% (43/74) of pedicle heights were larger than 3.5 mm. For pedicle width of subaxial spine, 75.7% (C3), 73% (C4), 82.4% (C5), 89.2% (C6), and 98.1% (C7, 1/54) were greater than 3.5 mm. Mean lamina width of subaxial cervical spine was 3.1 (C3), 2.7 (C4), 2.9 (C5), 3.8 (C6), and 4.0 mm (C7), respectively. Only 34.6% (127/370) of subaxial (C3-7) lamina thickness were greater than 3.5 mm. Mean length of lateral mass for the lateral mass screw placement was 9.28 (C3), 9.08 (C4), 8.81 (C5), 8.98 (C6), and 10.38 mm (C7). Conclusion : C1 lateral mass fixation could be limited by the morphometrics of lateral mass height. C2 translamina approach is preferable to C2 pedicle screw fixation. In subaxial spines, pedicle screw placement was preferable to trans-lamina screw placement, except at C7.
효율적인 복합재난 대응을 위한 복합시나리오 개발 방안 (방사능 사고 + 화학사고 복합유형을 중심으로 : 울산지역)
정호진(Hojin Jung),은종화(Chonghwa Eun),문일(Il Moon) 한국위험물학회 2021 한국위험물학회지 Vol.9 No.2
This study aims to develop the multi-scenario plan for the efficient response to ‘complex-disaster.’ Looking at the disaster situations that occur in our society, it shows a complex aspect in which several types of disasters occur simultaneously as subsequent disasters are induced from the underlying disasters that have a leading influence. Although it is necessary to prepare meticulously prepared countermeasures according to various disaster situations, the current understanding of ‘complex-disaster’ situation, as well as response plans for each type of disaster, is insufficient. In this context, this paper proposes a development theory for developing scenarios to for response to complex ‘complex-disasters’ based on chemical accident with radioactive disaster.
( Suyun Moon ),( Hwa-yong Lee ),( Donghwan Shim ),( Myungkil Kim ),( Kang-hyeon Ka ),( Rhim Ryoo ),( Han-gyu Ko ),( Chang-duck Koo ),( Jong-wook Chung ),( Hojin Ryu ) 한국균학회 2017 Mycobiology Vol.45 No.2
Sixteen genomic DNA simple sequence repeat (SSR) markers of Lentinula edodes were developed from 205 SSR motifs present in 46.1-Mb long L.edodes genome sequences. The number of alleles ranged from 3-14 and the major allele frequency was distributed from 0.17-0.96. The values of observed and expected heterozygosity ranged from 0.00-0.76 and 0.07-0.90, respectively. The polymorphic information content value ranged from 0.07-0.89. A dendrogram, based on 16 SSR markers clustered by the paired hierarchical clustering` method, showed that 33 shiitake cultivars could be divided into three major groups and successfully identified. These SSR markers will contribute to the efficient breeding of this species by providing diversity in shiitake varieties. Furthermore, the genomic information covered by the markers can provide a valuable resource for genetic linkage map construction, molecular mapping, and marker-assisted selection in the shiitake mushroom.