• 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 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.

• 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 Ginseng Research
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• v.37 no.1
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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.

• KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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• v.29 no.1
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• pp.43-49
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• 2023

This study reports on the preparation of sodium caseinate-cardanol (CasNa/CL)-based papers coated with different amounts of bentonite (BN) for use as a sustainable packaging material. Their chemical and morphological structures, mechanical properties, water vapor permeability, surface properties, and antioxidant activity of coated papers was assessed as a function of the BN content. The drying of the CasNa/CL coated papers led to the formation of pinholes on their surfaces owing to the presence of trapped water resulting from the difference in the drying rate between the external surface and the inside of the coated layers. Increasing the BN content reduced the pinholes on surface of CasNa/CL/BN coated papers and highly decreased the water vapor transmittance rate of the papers from 387.3±1.9 g/m²·day to 269.25±4.5 g/m²·day. Free radical scavenging assays indicated the addition of CL to the CasNa exhibited the antioxidant activity and antioxidant activity of CasNa/CL/BN did not changed as increase of BN contents. The improved water vapor barrier property and antioxidant activity of CasNa/CL/BN coated papers can be promised for various packaging applications.

• 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.

• The Journal of Korean Institute of Communications and Information Sciences
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• v.39C no.11
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• pp.1050-1067
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• 2014

CAS(Close Air Support) is aircraft attack against hostile targets that are in close proximity to friendly forces. Immediate CAS is the mission that attack unplanned targets, and especially the distribution of suitable aircraft assets makes huge effect on the result of immediate CAS mission. But It is hard to find a previous studies on immediate CAS sortie distribution with aircraft suitability. This study suggests a methodology with aircraft suitability for immediate CAS sortie distribution. The methodology consists of 3 steps. Firstly, we analyze target information for situational awareness. Secondly, we calculate each aircraft's suitability value per each target based on the result of previous analysis. Lastly, we suggest immediate CAS sortie distribution based on the aircraft adoptability value to a decision maker. This methodology will provide not only quantitative analysis, but also decision making of immediate CAS sortie distribution more timely and effectively.

• Korean Journal of Remote Sensing
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• v.36 no.5_2
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• pp.861-866
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• 2020

CAS(Compact Advanced Satellite) 500-1 satellite and its follow-up, CAS 500-2, are scheduled to be launched in 2021. For these satellites, a research project on 'CAS 500-1/2 Image Acquisition and Utilization Technology Development' has been carried out. This paper summarizes publications carried out under the project, papers presented within this special issue and contributions of the project.