• BMB Reports
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• v.57 no.1
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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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• v.54 no.5
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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 Life Science
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• v.30 no.3
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• pp.313-319
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• 2020

Recently, cattle epidemic diseases are caused by a pathogen such as a virus or bacterium. Such diseases can spread through various pathways, such as feed intake, respiration, and contact between livestock. Diagnosis based on the ELISA (Enzyme-linked immunosorbent assay) and PCR (Polymerase chain reaction) methods has limitations because these traditional diagnostic methods are time consuming assays that require multiple steps and dedicated equipment. In this review, we propose the use of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) Cas system based on DNA and RNA levels for early point-of-care diagnosis in cattle. In the CRISPR/Cas system, Cas effectors are classified into two classes and six subtypes. The Cas effectors included in class 2 are typically Cas9 in type II, Cas12 in type V (Cas12a and Cas12b) and Cas13 in type VI (Cas13a and Cas13b). The CRISPR/Cas system uses reporter molecules that are attached to the ssDNA strands. When the Cas enzyme cuts the ssDNA, these reporters either fluoresce or change color, indicating the presence of a specific disease marker. There are several steps in the development of a CRISPR/Cas system. The first is to select the Cas enzyme depending on DNA or RNA from pathogens (viruses or bacteria). Based on that, the next step is to integrate the optimal amplification, transducing method, and signal reporter. The CRISPR/Cas system is a powerful diagnostic tool using a gene-editing method, which is faster, better, and cheaper than traditional methods. This system could be used for early diagnosis of epidemic cattle diseases and help to control their spread.

• School Mathematics
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• v.10 no.2
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• pp.297-317
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• 2008

This study was designed to gain insight to adopt CAS into secondary level algebra education in Korea. Most inactive usage of calculators in math and most negative effects of calculators on their achievements of Korean students were shown in International studies such as TIMSS-R. A comparative study was carried out with consideration of mathematical backgrounds and technological environments. 8 Korean students and 26 US students in Grade 11 were participated in this study. Subjects' Problem solving process and their strategies of CAS usage in classical Box-problem with CAS were analyzed. CAS helped modeling by providing symbolic manipulation commands and graphs with students' mathematical knowledge. Results indicates that CAS requires shifts focus in algebraic contents: recognition of decimal & algebraic presentations of numbers; linking various presentations, etc. The extent of instrumentation effects on the selection of problem solving strategies among Korea and US students. Instrumentation

• Food Science and Biotechnology
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• v.17 no.1
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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.

• BMB Reports
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• v.56 no.2
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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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• v.12 no.3_4
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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 Arbitration Studies
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• v.31 no.2
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• pp.23-46
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• 2021

Sports disputes have specific characteristics compared to disputes that arise in the field of commerce. One particularity is the judicial system in which the CAS plays a key role as the International Supreme Court for sports-related matters. The CAS Code applies whenever the parties agree to submit a sports-related dispute to the CAS(Art. R27). Once the parties to the arbitration agreement have decided that the CAS Code should govern their proceedings. The parties' autonomy is, however, limited to the provisions of the CAS Code that provide for such a corresponding autonomy. The application of the mandatory rules contained in the CAS Code cannot be excluded. In CAS appeals arbitration proceedings, the Panel shall decide the dispute according to the applicable sports regulations and, subsidiarily, to the rules of law chosen by the parties(Art. R58). In international sports disputes, the uniform application and interpretation of the relevant regulations are essential. Therefore, Art. R58 should be applied as a mandatory rule without any changes. Regulations of the sports organizations are to be qualified as valid rules of law. CAS panels may also apply the so-called lex sportiva to the merits before considering statutory provisions of national jurisdictions. In this way, the specificities in (international) sports disputes can be taken into account without the need to further examine the application of national legal standards.

• 한국전산유체공학회:학술대회논문집
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• 2008.08a
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• pp.34.2-34.2
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• 2008

• Journal of Microbiology and Biotechnology
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• v.33 no.9
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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.