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Efficient CRISPR-Cas12f1-Mediated Multiplex Bacterial Genome Editing via Low-Temperature Recovery
( Se Ra Lim ),( Hyun Ju Kim ),( Sang Jun Lee ) 한국미생물생명공학회 2024 Journal of microbiology and biotechnology Vol.34 No.7
CRISPR-Cas system is being used as a powerful genome editing tool with developments focused on enhancing its efficiency and accuracy. Recently, the miniature CRISPR-Cas12f1 system, which is small enough to be easily loaded onto various vectors for cellular delivery, has gained attention. In this study, we explored the influence of temperature conditions on multiplex genome editing using CRISPR-Cas12f1 in an Escherichia coli model. It was revealed that when two distinct targets in the genome are edited simultaneously, the editing efficiency can be enhanced by allowing cells to recover at a reduced temperature during the editing process. Additionally, employing 3`-end truncated sgRNAs facilitated the simultaneous single-nucleotide level editing of three targets. Our results underscore the potential of optimizing recovery temperature and sgRNA design protocols in developing more effective and precise strategies for multiplex genome editing across various organisms.
Association between hair mineral and age, BMI and nutrient intakes among Korean female adults
Se Ra Hong,Seung Min Lee,Na Ri Lim,Hwan Wook Chung,Hong Seok Ahn 한국영양학회 2009 Nutrition Research and Practice Vol.3 No.3
This study was performed to investigate the association between hair mineral levels and nutrient intakes, age, and BMI in female adults who visited a woman's clinic located in Seoul. Dietary intakes were assessed by food frequency questionnaire and mineral levels were measured in collected hairs, and the relationship between these was examined. The average daily nutrient intakes of subjects were compared to those of the KDRIs, and the energy intake status was fair. The average intake of calcium in women of 50 years and over was 91.35% of KDRIs and the potassium intake was greatly below the recommended levels in all age groups. In the average hair mineral contents in subjects, calcium and copper exceeded far more than the reference range while selenium was very low with 85.19% of subjects being lower than the reference value. In addition, the concentrations of sodium, potassium, iron, and manganese in the hair were below the reference ranges in over 15% of subjects. The concentrations of sodium, chromium, sulfur, and cadmium in the hair showed positive correlations (P<0.05) with age, but the hair zinc level showed a negative correlation (P<0.05) with age. The concentrations of sodium, potassium, chromium, and cadmium in the hair showed positive correlations (P<0.05) with BMI. Some mineral levels in subjects of this study showed significant correlations with nutrient intakes, but it seems that the hair mineral content is not directly influenced by each mineral intake. As described above, some hair mineral levels in female adults deviated from the normal range, and it is considered that nutritional intervention to control the imbalance of mineral nutrition is required. Also, as some correlations were shown between hair mineral levels and age, BMI, and nutrient intakes, the possibility of utilizing hair mineral analysis for specific purposes in the future is suggested.
P-21 Introduction of national tuberculosis molecular typing service
( Se-mi Jeon ),( Na-ra Lim ),( Young-mi Kim ),( Dong-hyeok Kim ),( Seong-han Kim ),( Mi-sun Park ) 대한결핵 및 호흡기학회 2016 대한결핵 및 호흡기학회 추계학술대회 초록집 Vol.121 No.-
Molecular typing of Mycobacterium tuberculosis provides i) understanding the molecular epidemiology of nationwide TB, ii) early recognition of TB, iii) identification of incorrect TB diagnoses based on false-positive culture results, and iv) distinguishment relapse and re-infection. To achieve these objectives, national TB molecular typing service in many countries has been operated, such as TB Genotypes Information Management System of US CDC, TB national strain typing service of UK PHE, Molecular surveillance of MDR-TB in Europe. In 2014, Korea NRIH initiated TB molecular typing service to assist identify TB patients potentially involved in the same chain of recent TB transmission following routine contact investigations. A total of 1,788 isolates including epidemiological related strains were analyzed with 24 loci MIRU-VNTR and Spoligotyping and we established the database that was digitalization and enumeration, namely TBG type. The major lineage type of TB in our database is Beijing (81.04%). Results of TBG type showed that 51% of TBG type were unique pattern and the major TBG type was TBG0381 that take possession of 2.46%. Until now, national tuberculosis molecular typing service in Korea was limited to identify the suspected relationships between TB outbreaks cases, we will expand the service to distinguish relapse and re-infection in recurrent TB patient.
Lim, Sooman,Cho, Byungjin,Bae, Jaehyun,Kim, Ah Ra,Lee, Kyu Hwan,Kim, Se Hyun,Hahm, Myung Gwan,Nam, Jaewook IOP 2016 Nanotechnology Vol.27 No.43
<P>Scalable sub-micrometer molybdenum disulfide (<img ALIGN='MIDDLE' ALT='${\mathrm{MoS}}_{2}$' SRC='http://ej.iop.org/images/0957-4484/27/43/435501/nanoaa3ca9ieqn1.gif'/>) flake films with highly uniform coverage were created using a systematic approach. An electrohydrodynamic (EHD) printing process realized a remarkably uniform distribution of exfoliated <img ALIGN='MIDDLE' ALT='${\mathrm{MoS}}_{2}$' SRC='http://ej.iop.org/images/0957-4484/27/43/435501/nanoaa3ca9ieqn2.gif'/> flakes on desired substrates. In combination with a fast evaporating dispersion medium and an optimal choice of operating parameters, the EHD printing can produce a film rapidly on a substrate without excessive agglomeration or cluster formation, which can be problems in previously reported liquid-based continuous film methods. The printing of exfoliated <img ALIGN='MIDDLE' ALT='${\mathrm{MoS}}_{2}$' SRC='http://ej.iop.org/images/0957-4484/27/43/435501/nanoaa3ca9ieqn3.gif'/> flakes enabled the fabrication of a gas sensor with high performance and reproducibility for <img ALIGN='MIDDLE' ALT='${\mathrm{NO}}_{2}$' SRC='http://ej.iop.org/images/0957-4484/27/43/435501/nanoaa3ca9ieqn4.gif'/> and <img ALIGN='MIDDLE' ALT='${\mathrm{NH}}_{3}$' SRC='http://ej.iop.org/images/0957-4484/27/43/435501/nanoaa3ca9ieqn5.gif'/>.</P>
Se Ra Park,Jeong-Hwan Lee,Sohee Lim,Taek Min Kim,Seung Ho Lee,Dae Heon Kim,Chao Gao,Richard D. Cummings,Kisung Ko 한국당과학회 2021 한국당과학회 학술대회 Vol.2021 No.07
Overexpression of human epidermal growth factor receptor type 2 (HER2) protein is one of the prognostic factors for breast cancer, and HER2 is most likely to be recurrence when cancer metastasizes to lymph nodes. This study demonstrated the expression of the anti-HER2 camelid single domain antibody VHH in tobacco plant. And we confirmed the function of recombinant VHH for the treatment of breast cancer. The anti-HER2 VHH was fused to human IgG Fc domain along with KDEL endoplasmic reticulum (ER) (VHH-FcK) in a gene expression cassette. The plant gene expression cassette was transferred to tobacco plants using Agrobacterium-mediated transformation. The transformants were screened by PCR and western blots. ELISA confirmed the binding activity of purified anti-HER2 VHH-FcK to HER2 positive breast cancer cell line SK-BR-3. In conclusion, this study verified the possibility of anti-HER2 VHH-FcK as a therapeutic agent and suggests that the anti-breast cancer anti-HER2 VHH-FcK can be expressed, properly assembled and purified from plant expression system, which can be as an alternative system for antibody production. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT)(2021R1F1A1063869).
Electroacupuncture for Carpal Tunnel Syndrome: A Review of Randomized Controlled Trials
박철우,Lim Min Ji,Lee Se Won,Yi Yeon Hoo,Song Da Woon,Yu Sang Gu,Kim Min Ju,Oh Da Yoon,Choi Hyo Jung,Ju Ah Ra 대한침구의학회 2022 대한침구의학회지 Vol.39 No.2
This study aimed to examine the clinical efficacy of electroacupuncture treatment for carpal tunnel syndrome by reviewing published randomized controlled trials. Among the 186 studies retrieved from 7 online databases (PubMed, Cochrane library, CNKI, NDSL, RISS, OASIS, KMbase) on October 29, 2021, 4 studies were selected according to the inclusion, exclusion criteria, and were evaluated using risk of bias. Control groups for electroacupuncture were wearing a splint at night, traditional acupuncture, and medication. Methods such as total effective, functional status scale, symptom severity scale, electromyography, tip pinch strength, visual analogue scale, numeric rating scale, and ultrasound were used to evaluate the therapeutic effect. Electroacupuncture was reported to have significant treatment results compared with the control group in methods such as total effectiveness, electromyography, and tip pinch strength. However, the quality of the studies (using risk of bias) does not allow reliable conclusions to be made. Many high quality (low risk of bias) randomized controlled trials are needed to examine the efficacy of electroacupuncture treatment for carpal tunnel syndrome.
Park Se Ra,Ko Kisung,Lim Sohee,Cha So Yeon,정현주,Park Soon Ju,Myung Soon‐Chul,Kim Mi Kyung 한국곤충학회 2020 Entomological Research Vol.50 No.4
The monoclonal antibody (mAb) CO17-1A specifically binds to the tumor associated cell surface glycoprotein GA733 in colorectal cancer cells. Thus, mAb CO17-1A has the potential to act as an immune therapeutic protein against colorectal cancer. Recently, it was shown that the baculovirus insect cell expression system produces anti-colorectal cancer mAb CO17-1A. In this study, the colorectal cancer antibody mAb CO17-1A fused to the endoplasmic reticulum (ER) retention signal sequence (KDEL), and the (mAb CO17-1AK) was expressed in Spodoptera frugiperda Sf9 insect cells. The yield, cell cytotoxicity, and in vitro anti-tumor activity of mAb CO17-1AK were verified. Western blotting was performed to confirm that both heavy and light chains of mAb CO17-1A were expressed in Sf9 insect cells. The insect-derived mAb (mAbI ) CO17-1A was purified using a protein G affinity column. An in vitro wound healing assay was conducted to determine the inhibition activity of mAb CO17-1A during tumor cell migration, showing that mAbI CO17-1AK was effective as mammalian-derived mAb CO17-1A (mAbM CO17-1A). These results suggest that the insect cell expression system can produce and properly assemble mAbs that inhibit tumor cell migration.