• Title/Summary/Keyword: Knock-in

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Comparative Insecticidal Efficiency of Compounded Pyrethroids and Its Preparations Cooperated with the Several Synergists (Pyrethroid 살충제에 첨가되는 협력제에 따른 살충 효과)

  • 이숙경
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.189-195
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    • 1998
  • Comparative insecticidal efficiency between compounded pyrethroids and its preparations cooperated with several synergists was studied. As main components, Group A contained Permethrine.Phthalthrine, Group B contained Permethrine.Furameth rine and Group C contained Phthalthrine.Dichlorvos. As synergists Piperonyl Butoxide (P.B.O.), N-Octylbicycloheptene dicarboximide (MGK-264), Octachlorodipropylether (8-421) was cooperated in each group. These preparations was treated to either Blattella G. or Musca D. for comparing about insecticidal efficiency[Knock-down rate ($KT_{50},\;KT_{90}$) and cumulative mortaility(percent/hrs)]. When the synergists was added to each Group, the insecticidal efficiency was improved. The most potent synergists was P.B.O. and the potency was decreased in the rank order of S-421>MGK-264. The more the concentration of synergists was increased, the more the insecticidal efficiency was improved. The rank order was $1:5{\geq}1:4>1.3$ in all three groups. The mortality (percent) according to time was similar to either $KT_{50}\;or\;KT_{90}$. Knock-down rate was appropriate parameter for the indicator about potency.

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Endotoxin-induced renal tolerance against ischemia and reperfusion injury is removed by iNOS, but not eNOS, gene-deletion

  • Kim, Jee-In;Jang, Hee-Seong;Park, Kwon-Moo
    • BMB Reports
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    • v.43 no.9
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    • pp.629-634
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    • 2010
  • Endotoxin including lipopolysaccharide (LPS) confers organ tolerance against subsequent challenge by ischemia and reperfusion (I/R) insult. The mechanisms underlying this powerful adaptive defense remain to be defined. Therefore, in this study we attempted to determine whether nitric oxide (NO) and its associated enzymes, inducible NOS (iNOS) and endothelial NOS (eNOS, a constitutive NOS), are associated with LPS-induced renal tolerance against I/R injury, using iNOS (iNOS knock-out) or eNOS (eNOS knock-out) gene-deleted mice. A systemic low dose of LPS pretreatment protected kidney against I/R injury. LPS treatment increased the activity and expression of iNOS, but not eNOS, in kidney tissue. LPS pretreatment in iNOS, but not eNOS, knock-out mice did not protect kidney against I/R injury. In conclusion, the kidney tolerance to I/R injury conferred by pretreatment with LPS is mediated by increased expression and activation of iNOS.

Knock-down of Type 2 Inositol 1,4,5-Trisphosphate Receptors using Adenovirus in Adult Ventricular Myocytes (아데노바이러스를 이용한 성체 심실 근세포 이노시톨 1,4,5-삼인산 수용체 제 2 아형의 발현 억제)

  • Son, Min-Jeong;Subedi, Krishna P.;Woo, Sun-Hee
    • YAKHAK HOEJI
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    • v.54 no.1
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    • pp.8-12
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    • 2010
  • Inositol 1,4,5-trisphosphate ($IP_3$) receptor ($IP_3R$)-mediated signaling pathway is involved in many cellular processes including fertilization, apoptosis and neuronal function. Although cardiac myocytes express the $IP_3R$, its pathophysiological role has not been clearly understood because of limited selectivity of currently available pharmacological blockers. In the present study we constructed shRNA-expressing adenovirus to knock-down the type 2 $IP_3R$ ($IP_3R2$), a major subtype in cardiac ventricular myocytes, and demonstrated that the virus successfully eliminated the expression and localization of the $IP_3R2$. These results may provide a reliable tool for probing pathophysiological roles of the $IP_3R2$ in isolated intact cardiac myocytes.

Knock Characteristic Analysis of Gasoline and LPG Homogeneous Charge Compression Ignition Engine (가솔린과 LPG 예혼합 압축 착화 엔진의 노킹 특성)

  • Yeom, Ki-Tae;Bae, Choong-Sik
    • Transactions of the Korean Society of Automotive Engineers
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    • v.15 no.3
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    • pp.54-62
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    • 2007
  • The knock characteristics in an engine were investigated under homogeneous charge compression ignition (HCCI) operation. Liquefied petroleum gas (LPG)and gasoline were used as fuels and injected at the intake port using port fuel injection equipment. Di-methyl ether (DME) was used as an ignition promoter and was injected directly into the cylinder near compression top dead center (TDC). A commercial variable valve timing device was used to control the volumetric efficiency and the amount of internal residual gas. Different intake valve timingsand fuel injection amounts were tested to verify the knock characteristics of the HCCI engine. The ringing intensity (RI) was used to define the intensity of knock according to the operating conditions. The RI of the LPG HCCI engine was lower than that of the gasoline HCCI engine at every experimental condition. The indicated mean effective pressure (IMEP) dropped when the RI was over 0.5 MW/m2and the maximum combustion pressure was over 6.5MPa. There was no significant relationship between RI and fuel type. The RI can be predicted by the crank angle degree (CAD) at 50 CA. Carbon monoxide (CO) and hydrocarbon (HC) emissions were minimized at high RI conditions. The shortest burn duration under low RI was effective in achieving low HC and CO emissions.

TALEN Constructs and Validation for Targeting of SETDB1 Genomic DNA (SETDB1 genomic DNA 를 표적하는 TALEN construct 제작 및 분석)

  • Noh, Hee-Jung;Kang, Yoonsung;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1269-1275
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    • 2014
  • TALEN is a newly developed gene engineering method to knock out specific genes. It contains a DNA binding domain and a Fok1 nuclease domain in the TALEN plasmid. Therefore, the engineered TALEN construct can bind to any region of genomic DNA and cut the target nucleotide, thereby inducing mutation. In this study, we constructed two TALEN constructs targeted to a protein initiation codon (DBEX2) or the 25th upstream region (DBPR25) to enable mRNA synthesis of SETDB1 HMTase. We performed the TALEN cloning in two steps. The first step was from module vectors to pFUS array vectors. We confirmed successful cloning with a colony PCR experiment and Esp31 restriction enzyme digestion, which resulted in a smear band and a 1 Kb insert band, respectively The second step of the cloning was from a pFUS array vector to a mammalian TALEN expression vector. The engineered TALEN construct was sequenced with specific primers in an expression vector. As expected, a specific array from the module vectors was shown in the sequencing analysis. The specific module sequences were regularly arrayed in every 100 bp, and SETDB1 expression totally disappeared in the TALEN-DBEX2 transfection. PCR amplification targeting of DBEX2 was performed, and the PCR product was digested with a T7E1 restriction enzyme. The expression of SETDB1 was down-regulated in the TALEN-DBPR25 transfection. Morphological changes were also observed in the two TALEN constructs with transfected HeLa cells. These results suggest that the engineered TALEN constructs in two strategic approaches are very useful to knock-out of the SETDB1 gene and to study gene function.

Buckling behavior of composite cylindrical shells with cutout considering geometric imperfection

  • Heidari-Rarani, M.;Kharratzadeh, M.
    • Steel and Composite Structures
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    • v.30 no.4
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    • pp.305-313
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    • 2019
  • Creating different cutout shapes in order to make doors and windows, reduce the structural weight or implement various mechanisms increases the likelihood of buckling in thin-walled structures. In this study, the effect of cutout shape and geometric imperfection (GI) is simultaneously investigated on the critical buckling load and knock-down factor (KDF) of composite cylindrical shells. The GI is modeled using single perturbation load approach (SPLA). First, in order to assess the finite element model, the critical buckling load of a composite shell without cutout obtained by SPLA is compared with the experimental results available in the literature. Then, the effect of different shapes of cutout such as circular, elliptic and square, and perturbation load imperfection (PLI) is investigated on the buckling behavior of cylindrical shells. Results show that the critical buckling load of a shell without cutout decreases by increasing the PLI, whereas increasing the PLI does not have a great impact on the critical buckling load in the presence of cutout imperfection. Increasing the cutout area reduces the effect of the PLI, which results in an increase in the KDF.

CRISPR and Target-Specific DNA Endonucleases for Efficient DNA Knock-in in Eukaryotic Genomes

  • Lee, Seung Hwan;Kim, Sunghyun;Hur, Junho K
    • Molecules and Cells
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    • v.41 no.11
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    • pp.943-952
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    • 2018
  • The discovery and mechanistic understanding of target-specific genome engineering technologies has led to extremely effective and specific genome editing in higher organisms. Target-specific genetic modification technology is expected to have a leading position in future gene therapy development, and has a ripple effect on various basic and applied studies. However, several problems remain and hinder efficient and specific editing of target genomic loci. The issues are particularly critical in precise targeted insertion of external DNA sequences into genomes. Here, we discuss some recent efforts to overcome such problems and present a perspective of future genome editing technologies.

Evolution of CRISPR towards accurate and efficient mammal genome engineering

  • Ryu, Seuk-Min;Hur, Junseok W;Kim, Kyoungmi
    • BMB Reports
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    • v.52 no.8
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    • pp.475-481
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    • 2019
  • The evolution of genome editing technology based on CRISPR (clustered regularly interspaced short palindromic repeats) system has led to a paradigm shift in biological research. CRISPR/Cas9-guide RNA complexes enable rapid and efficient genome editing in mammalian cells. This system induces double-stranded DNA breaks (DSBs) at target sites and most DNA breakages induce mutations as small insertions or deletions (indels) by non-homologous end joining (NHEJ) repair pathway. However, for more precise correction as knock-in or replacement of DNA base pairs, using the homology-directed repair (HDR) pathway is essential. Until now, many trials have greatly enhanced knock-in or substitution efficiency by increasing HDR efficiency, or newly developed methods such as Base Editors (BEs). However, accuracy remains unsatisfactory. In this review, we summarize studies to overcome the limitations of HDR using the CRISPR system and discuss future direction.

Identification of Differentially Expressed Genes in the Dicer 1 Knock-down Mouse Embryos using Microarray

  • Lee, Jae-Dal;Cui, Xiang-Shun
    • Reproductive and Developmental Biology
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    • v.32 no.4
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    • pp.229-235
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    • 2008
  • Silencing of Dicer1 by siRNA did not inhibit development up to the blastocyst stage, but decreased expression of selected transcription factors, including Oct-4, Sox2 and Nanog, suggesting that Dicer1 gene expression is associated with differentiation processes at the blastocyst stage (Cui et al., 2007). In order to get insights into genes which may be linked with microRNA system, we compared gene expression profiles in Gapdh and Dicer1 siRNA-microinjected blastocysts using the Applied Biosystem microarray technology. Our data showed that 397 and 737 out of 16354 genes were up- and down-regulated, respectively, following siRNA microinjection (p<0.05), including 24 up- and 28 down-regulated transcription factors. Identification of genes that are preferentially expressed at particular Dicer1 knock down embryos provides insights into the complex gene regulatory networks that drive differentiation processes in embryos at blastocyst stage.

A Study on the Presure Resonance with Combustion Chamber Geometry for a Spark Ignition Engine (스파크 점화기관의 연소실 형상에 따른 공진현상 해석에 관한 연구)

  • Park, Kyoung-Suk;Jang, Seok-Hyung
    • Proceedings of the KSME Conference
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    • 2001.06d
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    • pp.890-895
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    • 2001
  • Pressure resonance frequency that is caused in the combustion chamber can be interpreted to acoustic analysis. Until now the pressure resonance has been assumed and calculated to a disc type combustion chamber that neglected the combustion chamber height because the knock occurs near the TDC(top dead center). In this research FEM(fine element method) has been used to calculate the pressure resonance frequency inside the experimental engine combustion. The reduce error of the resonance frequency obtained by FEM has decreased about 50% compared to the calculation of Draper's equation. Due to the asymmetry in the shape of the combustion chamber that was neglected in Draper's equation we could find out that a new resonance frequency could be generated. To make the experimental results equal we could know that the speed of sound that satisfies Draper's equation was selected 13% higher than all the pent-roof type combustion considered.

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