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Gene editing platform in Cattle
Sang-Eun Hahn,Soo-Young Yum,Song-Jeon Lee,Choong-Il Lee,Hee-Soo Kim,Hyeong-Jong Kim,Woo-Jae Choi,Ji-Hyun Lee,Woo-Sung Lee,Goo Jang 한국수정란이식학회 2016 한국수정란이식학회 학술대회 Vol.2016 No.10
The CRISPR/Cas9 system is proved to be a powerful tool for knock-out and knock-in in various species. By introducing genetic materials of two components (Cas9 and small guide (sg) RNA) into cells or pronuclear of the fertilized embryo, gene editing occurs. Some studies reported that efficiency of gene editing would be increased as Cas9 was integrated into cells or animals since Cas9 is indispensable in the CRISPR/Cas9 system. Accordingly, the production of Cas9 expressing cattle may provide the broadly used gene editing platform in cattle. For this study, Cas9 and RFP genes were cloned into PiggyBac (PB) transposon system. PB-Cas9-RFP and transposase were microinjected into 1436 in vitro fertilized embryos and 241 blastocysts were formed. Blastocysts with RFP expression accounting for 14.1% of total formed blastocysts were selected and transferred into 5 recipient cow. After gestation periods, four transgenic cattle were delivered without any veterinary assistance. From a transgenic cattle, ear skin tissue was collected for primary culture. On those primary cells, sgRNAs in DNA form for various genes such as PRNP, RB1 and BLG were transfected as 2ug of sgRNA per 5x105 cells using Nucleo factor system (Neon®, invitrogen, program#16). As expected, every group of each sgRNA delivered was confirmed to be mutated by T7E1assay. Those data demonstrated that for the first time, transgenic cattle with Cas9 expression were born, grown up to date and will be avaluable resource for genome-editing in cattle. This work was supported by BK21PLUS Program for Creative Veterinary Science and Seoul Milk Coop (SNU550-20160004).
Achievements and lessons learned from the operation of KSTAR plasma control system upgrade
Hahn, Sang-hee,Penaflor, B.G.,Milne, P.G.,Bak, J.G.,Eidietis, N.W.,Han, H.,Hong, J.S.,Jeon, Y.M.,Johnson, R.D.,Kim, H.-S.,Kim, HeungSu,Kim, Y.J.,Kwon, G.I.,Lee, W.R.,Woo, M.H.,Sammuli, B.S.,Walker, M. Elsevier 2018 Fusion engineering and design Vol.130 No.-
<P><B>Abstract</B></P> <P>Results on the integration and the operation of the KSTAR plasma control system (PCS) upgrade are given. Real-time hardware, new realtime-capable operating system, and a brand-new data acquisition are assembled in order to extend the performance and compatibility with modern computer systems. The first full commissioning and eventual routine use performed in 2016 so that the system can now acquire more than 400 channels for more than 100 seconds of data with 5 kHz sampling. The performance test results are summarized, featuring In/Out streaming echo tests and the synchronization verifications. Examples of general performance improvements are demonstrated, and additional features added to the software are also described.</P>
Involvement of Spontaneously Formed Cyclic Nucleotides in Cat Gastric Muscle Relaxation
Sang-Soo Sim,Hye-Jung Baek,Duck-Joo Rhie,Shin-Hee Yoon,Sang June Hahn,Yang-Hyeok Jo,Myung-Suk Kim 대한생리학회-대한약리학회 1999 The Korean Journal of Physiology & Pharmacology Vol.3 No.3
<P> Muscle strips and muscle cells from cat stomach were used to investigate whether spontaneously formed cyclic nucleotides were involved in the inhibition of gastric smooth muscle contraction. A phosphodiesterase inhibitor, 3-isobutyl-1-methylxanthine (IBMX), increased the levels of both cyclic GMP (cGMP) and cyclic AMP (cAMP) in resting state cells, while decreasing acetylcholine-induced muscle contraction. Under the influence of IBMX, SQ22536, an adenylyl cyclase inhibitor and methylene blue, a guanylyl cyclase inhibitor completely blocked increases in cAMP and cGMP respectively, without any effect on contraction. However, the combination of SQ22536 and methylene blue completely blocked increases in both cAMP and cGMP levels and stimulated contractions markedly even in the presence of IBMX. Muscle contraction inhibitors such as isoprenaline, vasoactive intestinal polypeptide and sodium nitroprusside also appeared to increase cyclic nucleotide levels which decreased contraction. Which nucleotide increased the most was dependent on the agonist used. Therefore, irrespective of the cyclic nucleotide class, the spontaneous formation of cyclic nucleotides should be considered in evaluating the mechanism of gastric smooth muscle relaxation.
Open Channel Block of Kv3.1 Currents by Fluoxetine
Sung, Min Ji,Ahn, Hye Sook,Hahn, Sang June,Choi, Bok Hee The Japanese Pharmacological Society 2008 Journal of pharmacological sciences Vol.106 No.1
<P>The action of fluoxetine, a serotonin reuptake inhibitor, on the cloned neuronal rat Kv3.1 channels stably expressed in Chinese hamster ovary cells was investigated using the whole-cell patch-clamp technique. Fluoxetine reduced Kv3.1 whole-cell currents in a reversible, concentration-dependent manner, with an IC<SUB>50</SUB> value and a Hill coefficient of 13.4 μM and 1.4, respectively. Fluoxetine accelerated the decay rate of inactivation of Kv3.1 currents without modifying the kinetics of current activation. The inhibition increased steeply between 0 and +30 mV, which corresponded with the voltage range for channel opening. In the voltage range positive to +30 mV, inhibition displayed a weak voltage dependence, consistent with an electrical distance δ of 0.38. The binding (k<SUB>+1</SUB>) and dissociation (k<SUB>−</SUB><SUB>1</SUB>) rate constants for fluoxetine-induced block of Kv3.1 were 5.7 μM<SUP>−</SUP><SUP>1</SUP>s<SUP>−</SUP><SUP>1</SUP> and 53.5 s<SUP>−</SUP><SUP>1</SUP>, respectively. The theoretical K<SUB>D</SUB> value derived by k<SUB>−</SUB><SUB>1</SUB>/k<SUB>+1</SUB> yielded 9.3 μM. Fluoxetine did not affect the ion selectivity of Kv3.1. Fluoxetine slowed the deactivation time course, resulting in a tail crossover phenomenon when the tail currents, recorded in the presence and absence of fluoxetine, were superimposed. Inhibition of Kv3.1 by fluoxetine was use-dependent. The present results suggest that fluoxetine acts on Kv3.1 currents as an open-channel blocker.</P>