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>

Genome surgery using Cas9 ribonucleoproteins for the treatment of age-related macular degeneration

Kim, Kyoungmi,Park, Sung Wook,Kim, Jin Hyoung,Lee, Seung Hwan,Kim, Daesik,Koo, Taeyoung,Kim, Kwang-eun,Kim, Jeong Hun,Kim, Jin-Soo Cold Spring Harbor Laboratory 2017 Genome research Vol.27 No.3

<P>RNA-guided genome surgery using CRISPR-Cas9 nucleases has shown promise for the treatment of diverse genetic diseases. Yet, the potential of such nucleases for therapeutic applications in nongenetic diseases is largely unexplored. Here, we focus on age-related macular degeneration (AMD), a leading cause of blindness in adults, which is associated with retinal overexpression of, rather than mutations in, the VEGFA gene. Subretinal injection of preassembled, Vegfa gene-specific Cas9 ribonucleoproteins (RNPs) into the adult mouse eye gave rise to mutagenesis at the target site in the retinal pigment epithelium. Furthermore, Cas9 RNPs effectively reduced the area of laser-induced choroidal neovascularization (CNV) in a mouse model of AMD. Genome-wide profiling of Cas9 off-target effects via Digenome-seq showed that off-target mutations were rarely induced in the human genome. Because Cas9 RNPs can function immediately after in vivo delivery and are rapidly degraded by endogenous proteases, their activities are unlikely to be hampered by antibody-and cell-mediated adaptive immune systems. Our results demonstrate that in vivo genome editing with Cas9 RNPs has the potential for the local treatment for nongenetic degenerative diseases, expanding the scope of RNA-guided genome surgery to a new dimension.</P>

Genome-wide target specificities of CRISPR RNA-guided programmable deaminases

Kim, Daesik,Lim, Kayeong,Kim, Sang-Tae,Yoon, Sun-heui,Kim, Kyoungmi,Ryu, Seuk-Min,Kim, Jin-Soo Nature Publishing Group, a division of Macmillan P 2017 Nature biotechnology Vol.35 No.5

<P>Cas9-linked deaminases, also called base editors, enable targeted mutation of single nucleotides in eukaryotic genomes. However, their off-target activity is largely unknown. Here we modify digested-genome sequencing (Digenome-seq) to assess the specificity of a programmable deaminase composed of a Cas9 nickase (nCas9) and the deaminase APOBEC1 in the human genome. Genomic DNA is treated with the base editor and a mixture of DNA-modifying enzymes in vitro to produce DNA double-strand breaks (DSBs) at uracil-containing sites. Off-target sites are then computationally identified from whole genome sequencing data. Testing seven different single guide RNAs (sgRNAs), we find that the rAPOBEC1-nCas9 base editor is highly specific, inducing cytosine-to-uracil conversions at only 18 +/- 9 sites in the human genome for each sgRNA. Digenome-seq is sensitive enough to capture off-target sites with a substitution frequency of 0.1%. Notably, off-target sites of the base editors are often different from those of Cas9 alone, calling for independent assessment of their genome-wide specificities.</P>

김정은 시기의 북한 고등교육정책 변화 방향 분석

김경미(Kim, Kyoungmi),김안나(Kim, Anna) 연세대학교 교육연구소 2020 미래교육학연구 Vol.33 No.3

본 연구의 목적은 사회주의 강성국가라는 국가적 과제에 대응하기 위한 김정은 시대 북한의 고등교육정책의 변화전략이 무엇인지 탐색하는 데 있다. 이에 본 연구는 북한 교육문화성이 발행한 교육신문 2012년~2019년 기사 중에서 ‘대학’, ‘고등교육’을 키워드로 수집한 1,255편의 기사를 분석하였다. 주요 결과는 다음과 같다. 첫째, 북한은 세계 일류급 고등교육을 강조하며 연구중심대학으로의 변화를 도모하며 과학기술연구 강조와 지식경제 시대에 대응하기 위한 지식을 생산하고자 하였다. 둘째, 대학을 유형별로 특성화하여 종합대학 일원화 및 박사 후 교육체계 마련, 직업기술대학체계를 확립하였다. 셋째, 대학교육의 질 향상을 위하여 원격교육 실행 및 교원의 교수 방법의 변화를 요구하였다. 넷째, 대학기능의 수월성 향상을 위하여 대학행정 시스템 및 대학입학 원격시험체계를 마련하고 인프라구축을 강화하고 있다. 이러한 연구결과를 토대로 본 연구는 북한과 같은 독특한 정치체제에서 교육정책의 변화가 북한사회의 변화에 어떤 영향을 미칠지 논의하였다. This study examines reform in higher education under Kim Jong-un that was motivated and defined by the strong influence of North Korea’s socialist ideology. To this end, 1,255 news articles containing the keywords “college” and “higher education” were analyzed. These were sourced from the Education Newspaper published by North Korean Education between 2012 and 2019.The study s conclusions are summarized below. First, North Korea seeks to develop a world-class higher education system modeled on research-centered institutions which emphasize science and technology for the new knowledge economy. Second, North Korea seeks to unify colleges, universities, post-doctoral studies, and vocational technology university system. Third, changes in the implementation of remote education and teaching methods were requested to improve the quality of university education. Fourth, infrastructure for improving university administration and remote testing for university admission have been emphasized and implemented to facilitate learning. This study discussed how changes in North Korean higher education policy reflect the unique characteristic of the political system and will shape future developments in North Korean society.

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>

Highly efficient RNA-guided base editing in mouse embryos

Kyoungmi Kim 한국실험동물학회 2017 한국실험동물학회 학술발표대회 논문집 Vol.2017 No.8

Influence of Optimism, Social Support, and Spirituality on COVID-19 Stress in Christian Church Community

Kim Kyoungmi,Lim Hyun Ju,Moon Eunsoo,Moon Sung Il 대한신경정신의학회 2023 PSYCHIATRY INVESTIGATION Vol.20 No.2

Objective Optimism, social support, and spirituality can be important factors related to coronavirus disease-2019 (COVID-19) stress. However, studies investigating the influence and interplay of optimism, social support, and spirituality on COVID-19 simultaneously are still few. This study is aimed to explore the influence of optimism, social support, and spirituality on COVID-19 stress in the Christian church community.Methods A total 350 participants were included in this study. This study was cross-sectionally conducted by using an online survey on optimism, social support, spirituality, and COVID-19 stress that were measured by the Life Orientation Test-Revised (LOT-R), Multidimensional Scale of Perceived Social Support Scale (MSPSS), Spiritual Well-Being Scale (SWBS), and COVID-19 Stress Scale for Korean People (CSSK). The prediction models for COVID-19 stress were analyzed by using univariate and multiple linear regression.Results Based on the results of univariate linear regression, subjective feelings on income (p<0.001) and health status (p<0.001), LOTR (p<0.001), MSPSS (p=0.025), and SWBS (p<0.001) scores were significantly associated with COVID-19 stress. The multiple linear regression model with subjective feelings on income and health status and SWSB score was significant (p<0.001) and explained 17.7% of the variance (R²=0.177).Conclusion This study showed that subjective feeling on low income, those who had poor health status, lower optimism, lower perceived social support, and lower spirituality were significantly affected with COVID-19 stress. Especially, the model with subjective feelings on income and health status, and spirituality showed highly significant effects, despite the interaction with associated factors. To cope with unpredictable stressful situations like the COVID-19 pandemic, integrated interventions on psycho-socio-spiritual aspect are warranted.

Highly efficient RNA-guided base editing in mouse embryos

Kyoungmi Kim 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

남자대학생의 대학생활 적응, 학업 스트레스가 성 차별주의에 미치는 영향

김경미(Kim Kyoungmi) 학습자중심교과교육학회 2018 학습자중심교과교육연구 Vol.18 No.23

본 연구는 남자 대학생의 대학생활 적응, 학업스트레스, 성 차별주의와의 관계를 파악하여 성 차별주의에 영향을 미치는 요인을 파악하고자 시도되었다. Y군에 위치한 4년제 대학의 남자대학생 195명을 대상으로, 2018년 3월부터 6월까지 자가 보고 식 설문조사로 시행되었다. 수집된 자료는 평균, 표준편차, Pearson correlation coefficient, stepwise multiple regression를 SPSS 19.0 프로그램을 이용하여 분석하였다. 연구결과 남자 대학생은 학과의 성비 균형과 불균형과 관계없이 대학생활적응이 낮을수록 학업스트레스와 성 차별주의가 높고, 학업스트레스가 높을수록 성 차별 주의가 높은 것으로 나타났으며, 학업스트레스와 온정적 성 차별주의는 남자대학생의 적대적 성 차별주의에 영향을 미치는 변수로 모형의 설명력은 42%로 나타났다. 따라서 학과의 남녀 성비에 따른 성 차별주의는 상관관계가 없으며, 대학생활 적응을 잘 할수록 학업에 대한 스트레스도 감소하고 성 차별주의도 감소할 것으로 사료된다. 특히 남자대학생들이 군 제대 후 복학하는 경우 대학생활에 적응할 수 있는 맞춤식 적응 프로그램이 필요하다. 또한 여자 대학생을 단순한 경쟁의 대상이 아닌 동료로 인식을 전환할 수 있는 계기를 마련해 줘야 한다. 이를 위해 다양한 교육과정의 개선 및 비교과활동을 개발하여 남자대학생의 적극 참여를 권장해야할 것이다. Purpose: This study was attempted to identify the factors influencing sexism by identifying the relation between male college students’ adaptation to college life, academic stress, and sexism. Method: Self-reported survey was conducted on 195 male students of four-year-course colleges located in Y County from March to June 2018. Analysis of the collected data was conducted using SPSS 19.0 program, and descriptive statistics, Pearson correlation coefficient, stepwise multiple regression. Results: It appeared that male students had high academic stress and sexism when their adaptation to college life was lower regardless of the gender balance and imbalance of their department, and sexism turned out to be high when academic stress was higher. Academic stress and compassionate sexism were variables influencing the hostile sexism of male students, and the explanation power of the model was 42%. Therefore, sexism according to the ratio of men and women of the department had no correlation, and it is considered that the better the student adapts to college life, the lower the stress on academics and sexism. Conclusion: in case of male college students returning to school after being discharged from the military, a customized adaptation program that enables them to adapt to college life is needed. Furthermore, an opportunity should be provided for the male students to change their perception of female students as their colleagues rather than their competitors. For this purpose, active participation of male students should be encouraged by improving various curriculums and developing non-subject activities.

대학생의 따돌림 경험과 우울의 관계에서 감사와 삶의 의미의 매개효과

김경미(Kyoungmi Kim) 한국동서정신과학회 2022 동서정신과학 Vol.25 No.2

본 연구는 대학생의 따돌림 경험이 우울에 어떠한 영향을 미치는지 그리고 따돌림 경험이 우울에 영향을 미치는 과정에서감사와 삶의 의미가 매개하는지 알아보고자 하였다. 이를 위하여 경상도와 전라도에 소재한 4년제 대학교에서 대학생330명에게 따돌림 경험, 우울, 감사, 삶의 의미에 관한 설문을 실시하여 분석하였다. 이중매개효과를 검증하기 위해 기술통계 및 상관분석, SPSS Macro 부트스트래핑(bootstrapping) 방법을 적용하였다. 연구결과, 첫째, 대학생의 따돌림 경험이 높을수록 우울이 높았다. 둘째, 대학생의 따돌림 경험과 우울의 관계에서 감사가 부분매개하였다. 셋째, 대학생의 따돌림 경험과 우울의 관계에서 삶의 의미의 매개효과는 없었다. 넷째, 대학생의 따돌림 경험은 감사와 삶의 의미의 이중매개 경로를 거쳐 우울에 영향을 미쳤다. 이러한 결과를 중심으로 대학생의 따돌림 경험이 우울에 미치는 영향을 감소시키기 위해서는 감사와 삶의 의미의 보호요인이 중요함을 논의하였다. The purpose of this study was to examine the double mediating effects of gratitude and meaning in life on the relationships between bullying experience and depression in college students. For this study, 330 college students were surved on bullying experience, depression(CESD), gratitude, and meaning in life at four-year universities located in Gyeongsang-do and Jeolla-do. Descriptive statistics, correlation analysis, regression, SPSS Macros, and bootstrapping methods were applied to verify the double mediating effects. The results of this study were as follows. First, the higher the bullying experience of college students, the higher the depression. Second, gratitude partially mediated in the relationship between bullying experience and depression of college students. Third, there was no mediating effect of the meaning of life in the relationship between bullying experience and depression of college students. Lastly, the bullying experience indirectly influenced depression through the double mediation effect of gratitude and meaning in life. Focusing on these results, it was discussed that the protective factors of gratitude and meaning of life are important to reduce the effects of college students' bullying experiences on depression.