• BMB Reports
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• 제57권1호
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• pp.40-49
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• 2024

Prokaryotes encode clustered regularly interspaced short palindromic repeat (CRISPR) arrays and CRISPR-associated (Cas) genes as an adaptive immune machinery. CRISPR-Cas systems effectively protect hosts from the invasion of foreign enemies, such as bacteriophages and plasmids. During a process called 'adaptation', non-self-nucleic acid fragments are acquired as spacers between repeats in the host CRISPR array, to establish immunological memory. The highly conserved Cas1-Cas2 complexes function as molecular recorders to integrate spacers in a time course manner, which can subsequently be expressed as crRNAs complexed with Cas effector proteins for the RNA-guided interference pathways. In some of the RNA-targeting type III systems, Cas1 proteins are fused with reverse transcriptase (RT), indicating that RT-Cas1-Cas2 complexes can acquire RNA transcripts for spacer acquisition. In this review, we summarize current studies that focus on the molecular structure and function of the RT-fused Cas1-Cas2 integrase, and its potential applications as a directional RNA-recording tool in cells. Furthermore, we highlight outstanding questions for RT-Cas1-Cas2 studies and future directions for RNA-recording CRISPR technologies.

• Nuclear Engineering and Technology
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• 제54권5호
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• pp.1535-1540
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• 2022

A solid tungsten target was used at the China Spallation Neutron Source (CSNS) with 100 kW proton beam power. To improve the lifetime, hot isostatic pressing (HIP) process was selected to bond tantalum cladding with tungsten plates. Radioactive isotope ¹⁸²Ta, an activation product of tantalum, was found in the cooling water after a period of operation, however, no radioactive isotopes of ¹⁸⁷W was found, which shows the tantalum layer remained mostly intact. The CSNS Target-1 had been operating safely for three years and was replaced by Target-2 in August 2020.

• Journal of Microbiology and Biotechnology
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• 제33권9호
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• pp.1228-1237
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• 2023

The CRISPR-Cas system has emerged as the most efficient genome editing technique for a wide range of cells. Delivery of the Cas9-sgRNA ribonucleoprotein complex (Cas9 RNP) has gained popularity. The objective of this study was to develop a quantitative polymerase chain reaction (qPCR)-based assay to quantify the double-strand break reaction mediated by Cas9 RNP. To accomplish this, the dextransucrase gene (dsr) from Leuconostoc citreum was selected as the target DNA. The Cas9 protein was produced using recombinant Escherichia coli BL21, and two sgRNAs were synthesized through in vitro transcription to facilitate binding with the dsr gene. Under optimized in vitro conditions, the 2.6 kb dsr DNA was specifically cleaved into 1.1 and 1.5 kb fragments by both Cas9-sgRNA365 and Cas9-sgRNA433. By monitoring changes in dsr concentration using qPCR, the endonuclease activities of the two Cas9 RNPs were measured, and their efficiencies were compared. Specifically, the specific activities of dsr365RNP and dsr433RNP were 28.74 and 34.48 (unit/㎍ RNP), respectively. The versatility of this method was also verified using different target genes, uracil phosphoribosyl transferase (upp) gene, of Bifidobacterium bifidum and specific sgRNAs. The assay method was also utilized to determine the impact of high electrical field on Cas9 RNP activity during an efficient electroporation process. Overall, the results demonstrated that the qPCR-based method is an effective tool for measuring the endonuclease activity of Cas9 RNP.

• BMB Reports
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• 제56권2호
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• pp.102-107
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• 2023

Genome editing using CRISPR-associated technology is widely used to modify the genomes rapidly and efficiently on specific DNA double-strand breaks (DSBs) induced by Cas9 endonuclease. However, despite swift advance in Cas9 engineering, structural basis of Cas9-recognition and cleavage complex remains unclear. Proper assembly of this complex correlates to effective Cas9 activity, leading to high efficacy of genome editing events. Here, we develop a CRISPR/Cas9-RAD51 plasmid constitutively expressing RAD51, which can bind to single-stranded DNA for DSB repair. We show that the efficiency of CRISPR-mediated genome editing can be significantly improved by expressing RAD51, responsible for DSB repair via homologous recombination (HR), in both gene knock-out and knock-in processes. In cells with CRISPR/Cas9-RAD51 plasmid, expression of the target genes (cohesin SMC3 and GAPDH) was reduced by more than 1.9-fold compared to the CRISPR/Cas9 plasmid for knock-out of genes. Furthermore, CRISPR/Cas9-RAD51 enhanced the knock-in efficiency of DsRed donor DNA. Thus, the CRISPR/Cas9-RAD51 system is useful for applications requiring precise and efficient genome edits not accessible to HR-deficient cell genome editing and for developing CRISPR/Cas9-mediated knockout technology.

• Environmental Analysis Health and Toxicology
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• 제12권3_4호
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• pp.1-13
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• 1997

First- and second-order kinetic oxidation rates of trichlorobenzenes (TCBs) were obtained and compared by a chemical activation system (CAS) which mimics mixed functional oxidase activity. The system consists of EDTA, ferrous sulfate, ascorbic acid, and $H_2O_2$ in potassium phosphdte buffer (monobasic at pH 7.4). The rate of transformation in CAS was enhanced in the presence and absence of catalase in the sequence 1, 2, 3-TCB < 1, 2, 4-TCB < 1, 3, 5-TCB. In general, the rates of degradation were greater in the test media with catalase. The effect of photolysis on the degradation of the TCBs with the CAS were examined. Sensitized photolysis with nitrite, Fenton's reagent, TiO$_2$ and triethylamine (TEA) studied in concert with the CAS demonstrated significant enhancement of the degradation rate of TCBs. Disappearance rates of TCBs in CAS with prior photolysis or prior photosensitization were at least 10-fold higher than the sum of the rate for each single experiment. This study proves that the combination of the CAS and photolysis can be used as a suitable technique for enhancing degradation of TCBs in aqueous systems.

• Journal of Ginseng Research
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• 제37권1호
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• pp.94-99
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• 2013

Proteinuric conditions demonstrate structural and compositional changes of the foot processes and slit diaphragms between podocytes. p130Cas in podocytes serves as an adapter protein anchoring glomerular basement membrane to actin filaments of podocyte cytoskeleton. To investigate the effect of ginseng total saponin (GTS) on the pathologic changes of podocyte p130Cas induced by diabetic conditions, we cultured mouse podocytes under: 1) normal glucose (5 mM, control); 2) high glucose (HG, 30 mM); 3) advanced glycosylation endproducts (AGE)-added; or 4) HG plus AGE-added conditions and treated with GTS. In confocal imaging, p130Cas colocalized with zonula occludens-1 and synaptopodin connecting to F-actin. However, diabetic conditions relocalized p130Cas molecules at perinuclear cytoplasmic area and reduced the intensity of p130Cas. In Western blotting, diabetic conditions, especially HG plus AGE-added condition, decreased cellular p130Cas protein levels at 24 and 48 h. GTS improved such quantitative and qualitative changes. These findings imply that HG and AGE have an influence on the redistribution and amount of p130Cas of podocytes, which can be reversed by GTS.

• 한국포장학회지
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• 제29권1호
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• pp.43-49
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• 2023

본 연구에서는 CasNa/CL의 물리적 특성을 개선하기 위하여 BN을 충진제로 활용하여 CasNa/CL/BN코팅제 및 코팅지들을 제조하였다. BN의 함량변화에 따라 제조한 CasNa/CL코팅지와 CasNa/CL/BN코팅지들의 화학적 및 형태학적 특성을 분석하였고, 기계적강도, 수증기차단특성, 표면특성, 항산화특성에 대한 분석을 통해 포장소재로써 적용 가능성을 확인하였다. SEM분석결과, CasNa/CL코팅지 표면에서 핀홀 현상이 발생하는 것을 확인하였다. 하지만, BN함량이 증가함에 따라 핀홀 현상은 감소되는 경향을 보였고 표면거칠기는 증가되는 경향을 확인할 수 있었다. BN 함량이 증가함에 따라 CasNa/CL/BN코팅지들의 연신률 및 수증기차단성이 개선되는 것을 확인할 수 있었다. BN함량 증가에도 불구하고 CasNa/CL/BN코팅지들의 항산화특성은 CasNa/CL코팅지와 유사한 경향을 보임을 확인하였다. 자연유래소재인 CasNa, CL 및 BN을 활용한 코팅지의 경우 친환경 포장소재로써 활용이 가능할 것으로 판단되지만, CL 및 BN과 CasNa와의 혼화성 및 분산성 개선 방안에 대한 추가적인 연구가 필요함을 확인하였다.

• Food Science and Biotechnology
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• 제17권1호
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• pp.8-14
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• 2008

The influence of salt concentration on the stability of sodium caseinate (CAS)-stabilized emulsions (20 wt% corn oil, 3.2 wt% CAS, 5 mM imidazole/acetate buffer, pH 7) was examined. In the absence of salt, laser diffraction measurements and optical microscopy measurements indicated there were some large oil droplets ($d>10\;{\mu}m$) in the emulsions stabilized by 0.8 to 3.2 wt% of CAS. The droplet aggregation (mostly droplet coalescence) observed in the emulsions containing ${\leq}2.8\;wt%$ CAS tended to decrease as the CAS concentration increased, however, after which concentration (at 3.2 wt% CAS) depletion flocculation occurred. The addition of $CaCl_2$ (5-20 mM) into the emulsions stabilized by 3.2 wt% CAS prevented the depletion flocculation although there was a small fraction of relatively large individual droplets in the emulsions, which was attributed to electrostatic screening effect and bridging effect of calcium ion. This study has shown that calcium ion that has been reputed to promote droplet aggregation could improve emulsion stability against droplet aggregation in CAS-stabilized emulsions.

• 한국통신학회논문지
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• 제39C권11호
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• pp.1050-1067
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• 2014

CAS는 근접항공지원 작전으로 아군과 근접해 있는 상황에서 적을 항공기로 공격하는 작전이다. CAS의 여러형태 중에 긴급 CAS는 CAS 임무 형태 중에 사전 계획 없이 요청한 표적에 대하여 공격하는 임무로, 적절한 항공기를 분배하는 것이 임무 결과에 커다란 영향을 미친다. 하지만 긴급 CAS 분배에 관련한 이전 연구에서는 항공기의 적합도를 고려한 경우를 찾기 힘들었다. 2014본 연구는 항공기 적합도를 고려한 긴급 CAS 자원 분배 방법론을 제시하고자 한다. 방법론은 총 3단계로 이루어져 있으며 1단계에서, 표적 정보를 바탕으로 상황분석을 실시하고, 2단계에서는, 상황분석 결과를 이용하여 각 표적별로 타격 자산들을 적합도를 정량적으로 산출하며, 3단계에서는 산출한 적합도를 바탕으로 산정한 CAS 분배 추천 안을 결정권자에게 제시한다. 이 방법론은 긴급 CAS 자산 분배에 관한 정량적인 분석을 제공함과 동시에, 추천 대안을 제시함으로써 의사결정을 보다 신속하고 효율적으로 할 수 있도록 지원한다.

• 대한원격탐사학회지
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• 제36권5_2호
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• pp.861-866
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• 2020

2021년에 발사예정인 국토관측위성의 처리와 활용을 위한 '국토위성정보 수집 및 활용기술개발' 과제가 2018년 7월부터 시작되어 2020년 12월에 종료예정이다. 본 논문에서는 그 동안 상기 과제를 수행하며 발표한 논문들과 본 특별호에 수록한 논문들을 소개하고 과제의 성과에 대해서 간략히 정리하고자 한다.