• Title/Summary/Keyword: SiRNA

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Gene Expression Profiling by RNA Sequencing in Mature/Immature Oocytes of Chicken (닭의 성숙/미성숙란에서 RNA Sequencing을 이용한 유전자 발현 양상 고찰)

  • Kang, Kyung-Soo;Jang, Hyun-Jun;Park, Mi Na;Choi, Jung-Woo;Chung, Won-Hyong;Heo, Kang-Nyeong;Choe, Chang-Yong;Kim, Young-Joo;Lee, Si-Woo;Cho, Eun-Seok;Kim, Namshin;Kim, Tae-Hun;Han, Jae-Yong;Lee, Kyung-Tai
    • Korean Journal of Poultry Science
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    • v.41 no.4
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    • pp.287-296
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    • 2014
  • Chicken eggs undergo various physiological changes during egg maturation. To study genes associated with the egg maturation in pre-ovulation (immature) and post-ovulation (mature), we compared gene expression patterns between in the immature egg and mature egg using RNA sequencing data. Mature and immature eggs were obtained from a Heuksaek Jaerae-jong of Korean native chicken. Total RNAs obtained from the eggs were sequenced by Illumina HiSeq 2000 platform, and the generated sequence reads were mapped to Galgal4 reference sequence assembly using Tuxedo Protocol. From the comparison of the RNA sequencing data, 315 genes were differentially expressed between mature and immature eggs, and 46 genes were only detected in immature egg. Further gene ontology (GO) analysis was performed for the differentially expressed genes using DAVID, showing that 29 and 28 GO terms were independently clustered from mature and immature, respectively. From those clustered GO terms, genes related to germ cell development, sex differentiation and defense response to bacterium were mainly expressed in the immature egg, while genes related to regulation of apoptosis, steroid metabolic process and lipid homeostasis were mainly detected in the mature egg. Our results could contribute to understand egg maturation before and after ovulation, and develop genetic markers for improving egg quality and productivity.

Effect of Atrazine, Perfluorooctanoic Acid and Zearalenone on IFNγ, TNFα, and IL-5 mRNA Expression in Jurkat Cells

  • Lee, Sung-Woo;Son, Hwa-Young;Yoon, Won-Kee;Jung, Ju-Young;Park, Bae-Keun;Cho, Eun-Sang;Park, Sang-Joon;Kim, Tae-Hwan;Ryu, Si-Yun
    • Biomolecules & Therapeutics
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    • v.18 no.3
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    • pp.286-293
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    • 2010
  • Cytokine production is a sensitive indicator for monitoring perturbations of the immune system by xenobiotics in animals and humans. In the present study, we evaluated the changes in $IFN{\gamma}$, IL-5 and $TNF{\alpha}$ mRNA expression after atrazine (ATZ), perfluorooctanoic acid (PFOA) or zearalenone (ZEA) exposure in Jurkat cells. The IC50 (concentration for a 50% inhibition of cell proliferation) of PFOA and ZEA after 3 days culture were $226.6\;{\mu}M$ and $52.6\;{\mu}M$, respectively. The effects of ATZ on cytokine expression followed in increasing order of $IFN{\gamma}$>IL-5>$TNF{\alpha}$ at $3\;{\mu}M$ and at the lower concentrations the degree of effects on three cytokines were less clear between the cytokines when compared to control level. PFOA had marked increasing effect in order of $IFN{\gamma}$>$TNF{\alpha}$>IL-5 mRNA expression at IC50, and these patterns were continued at the lower concentrations, IC50/2 and IC50/4. ZEA caused the overexpression of cytokine mRNAs in order of IL-5>$IFN{\gamma}$>$TNF{\alpha}$ at both IC50 and IC50/2, and at IC50/4 the overexpression order was IL-5>$TNF{\alpha}$. On other hand, $IFN{\gamma}$ was less distinct compared to the control. These data indicate that ATZ, PFOA and ZEA caused the overtranscription of $IFN{\gamma}$, IL-5 and $TNF{\alpha}$ mRNA, and the overproduction of these cytokines may eventually lead to immune disorders.

Knockdown of HMGN5 Expression by RNA Interference Induces Cell Cycle Arrest in Human Lung Cancer Cells

  • Chen, Peng;Wang, Xiu-Li;Ma, Zhong-Sen;Xu, Zhong;Jia, Bo;Ren, Jin;Hu, Yu-Xin;Zhang, Qing-Hua;Ma, Tian-Gang;Yan, Bing-Di;Yan, Qing-Zhu;Li, Yan-Lei;Li, Zhen;Yu, Jin-Yan;Gao, Rong;Fan, Na;Li, Bo;Yang, Jun-Ling
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3223-3228
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    • 2012
  • HMGN5 is a typical member of the HMGN (high mobility group nucleosome-binding protein) family which may function as a nucleosomal binding and transcriptional activating protein. Overexpression of HMGN5 has been observed in several human tumors but its role in tumorigenesis has not been fully clarified. To investigate its significance for human lung cancer progression, we successfully constructed a shRNA expression lentiviral vector in which sense and antisense sequences targeting the human HMGN5 were linked with a 9-nucleotide loop. Inhibitory effects of siRNA on endogenous HMGN5 gene expression and protein synthesis were demonstrated via real-time RT-PCR and western blotting. We found HMGN5 silencing to significantly inhibit A549 and H1299 cell proliferation assessed by MTT, BrdU incorporation and colony formation assays. Furthermore, flow cytometry analysis showed that specific knockdown of HMGN5 slowed down the cell cycle at the G0/G1 phase and decreased the populations of A549 and H1299 cells at the S and G2/M phases. Taken together, these results suggest that HMGN5 is directly involved in regulation cell proliferation in A549 and H1299 cells by influencing signaling pathways involved in cell cycle progression. Thus, our finding suggests that targeting HMGN5 may be an effective strategy for human lung cancer treatment.

MiR-26a promotes apoptosis of porcine granulosa cells by targeting the 3β-hydroxysteroid-Δ24-reductase gene

  • Zhang, Xiaodong;Tao, Qiangqiang;Shang, Jinnan;Xu, Yiliang;Zhang, Liang;Ma, Yingchun;Zhu, Weihua;Yang, Min;Ding, Yueyun;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.4
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    • pp.547-555
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    • 2020
  • Objective: Apoptosis of ovarian granulosa cells (GCs) affects mammalian follicular development and fecundity. This study aimed to explore the regulatory relationship between microRNA-26a (miR-26a) and the 3β-hydroxysteroid-Δ24-reductase gene (DHCR24) gene in porcine follicular granular cells (pGCs), and to provide empirical data for the development of methods to improve the reproductive capacity of pigs. Methods: The pGCs were transfected with miR-26a mimic, miR-26a inhibitor and DHCR24-siRNA in vitro. The cell apoptosis rate of pGCs was detected by the flow cytometry. The secretion levels of estradiol (E2) and progesterone (P) in pGCs were detected by enzyme-linked immunosorbent assay. Double luciferase validation system was used to detect the binding sites between miR-26a and DHCR24 3'-UTR region. Qualitative real-time polymerase chain reaction and Western blotting were used to verify the DHCR24 mRNA and protein expression in pGCs, respectively, after transfecting with miR-26a mimic and miR-26a inhibitor. Results: Results showed that enhancement of miR-26a promoted apoptosis, and inhibited E2 and P secretion in pGCs. Meanwhile, inhibition of DHCR24 also upregulated the Caspase-3 expression, reduced the BCL-2 expression, promoted pGCs apoptosis, and inhibited E2 and P secretion in pGCs. There were the binding sites of miR-26a located within DHCR24 3'-UTR. Up-regulation of miR-26a inhibited DHCR24 mRNA and protein expression in pGCs. Conclusion: This study demonstrates that miR-26a can promote cell apoptosis and inhibit E2 and P secretion by inhibiting the expression of DHCR24 in pGCs.

Postsurgical Wound Infection Caused by Mycobacterium conceptionense Identified by Sequencing of 16S rRNA, hsp65, and rpoB Genes in an Immunocompetent Patient (16S rRNA, hsp65, 및 rpoB 염기순서분석으로 동정한 Mycobacterium conceptionense에 의한 면역능이 정상인 환자에서 발생한 수술후 창상감염)

  • Lee, Ja Young;Kim, Si Hyun;Shin, Jeong Hwan;Lee, Hyun-Kyung;Lee, Young Min;Song, Sae Am;Bae, Il Kwon;Kim, Chang-Ki;Jun, Kyung Ran;Kim, Hye Ran;Lee, Jeong Nyeo;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.17 no.1
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    • pp.23-27
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    • 2014
  • Rapidly growing mycobacteria are ubiquitous in the environment and are increasingly being recognized as opportunistic pathogens. Recently, a new species, Mycobacteium conceptionense, has been validated from the Mycobacterium fortuitum third biovariant complex by molecular analysis. However, there are few reports, and postsurgical wound infection by this species is rare. We report a case of postsurgical wound infection caused by M. conceptionense in an immunocompetent patient that was identified by a sequencing analysis of 16S rRNA, hps65, and rpoB genes.

RNA viruses in Pleurotus ostreatus

  • Lee, Jung-Sun;Lee, Nam-Joo;Ha, Si-Jung;Kim, So-Yeon;Kang, Eun-Joo;Kang, Dong-Suk;Rho, Hyun-Su;Chang, Moo-Ung;Lee, Jong-Kyu;Lee, Kyung-Hee;Lee, Hyun-Sook
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2004.10a
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    • pp.46-46
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    • 2004
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HER2 induces expression of leptin in human breast epithelial cells

  • Cha, Yujin;Kang, Youjin;Moon, Aree
    • BMB Reports
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    • v.45 no.12
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    • pp.719-723
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    • 2012
  • A close association between the obesity hormone leptin and breast cancer progression has been suggested. The present study investigated the molecular mechanism for enhanced leptin expression in breast cancer cells and its functional significance in breast cancer aggressiveness. We examined whether leptin expression level is affected by the oncoprotein human epidermal growth factor receptor2 (HER2), which is overexpressed in ~30% of breast tumors. Here, we report, for the first time, that HER2 induces transcriptional activation of leptin in MCF10A human breast epithelial cells. We also showed that p38 mitogen-activated protein kinase signaling was involved in leptin expression induced by HER2. We showed a crucial role of leptin in the invasiveness of HER2-MCF10A cells using an siRNA molecule targeting leptin. Taken together, the results indicate a molecular link between HER2 and leptin, providing supporting evidence that leptin represents a target for breast cancer therapy.

Highly Efficient Encapsulation of Anionic Small Molecules in Asymmetric Liposome Particles

  • Lee, Myung Kyu
    • Applied Science and Convergence Technology
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    • v.24 no.6
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    • pp.284-288
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    • 2015
  • Anionic small molecules are hard to penetrate the cell membranes because of their negative charges. Encapsulation of small molecules into liposome particles can provide target specific delivery of them. In our previous study, siRNA could be efficiently encapsulated into liposome particles using an asymmetric preparation method of liposomes. In this study, the same method was applied for encapsulation of small anionic fluorescent chemicals such as calcein and indocyanine green (ICG). More than 90% fluorescent chemicals were encapsulated in the asymmetric liposome particles (ALPs). No intracellular fluorescent signal was observed in the tumor cells treated with the unmodified calcein/ALPs and ICG/ALPs, whereas the surface modification with a cell-penetrating polyarginine peptide (R8 or R12) allows cellular uptake of the ALPs. The results demonstrate that the ALPs encapsulating small anionic drugs will be useful for target-specific delivery after modification of target-specific ligands.

Functions of PIWI proteins in spermatogenesis

  • Bak, Chong-Won;Yoon, Tae-Ki;Choi, Young-Sok
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.2
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    • pp.61-67
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    • 2011
  • Recently, a significant understanding of the molecular mechanisms regulating spermatogenesis has been achieved utilizing small RNA molecules (small RNAs), including small interfering RNAs (siRNAs), microRNAs (miRNAs), and Piwi-interacting RNAs (piRNAs) which emerged as important regulators of gene expression at the post-transcriptional or translation level. piRNAs are only present in pachytene spermatocytes and round spermatids, whereas miRNAs are expressed abundantly in male germ cells throughout spermatogenesis. This review is aimed at providing a glimpse of piRNAs and their interacting family proteins such as PIWIL1, PIWIL2, and PIWIL4 in spermatogenesis.

Drug Discovery Perspectives of Antisense Oligonucleotides

  • Yeonjoon Kim
    • Biomolecules & Therapeutics
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    • v.31 no.3
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    • pp.241-252
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    • 2023
  • The era of innovative RNA therapies using antisense oligonucleotides (ASOs), siRNAs, and mRNAs is beginning. Since the emergence of the concept of ASOs in 1978, it took more than 20 years before they were developed into drugs for commercial use. Nine ASO drugs have been approved to date. However, they target only rare genetic diseases, and the number of chemistries and mechanisms of action of ASOs are limited. Nevertheless, ASOs are accepted as a powerful modality for next-generation medicines as they can theoretically target all disease-related RNAs, including (undruggable) protein-coding RNAs and non-coding RNAs. In addition, ASOs can not only downregulate but also upregulate gene expression through diverse mechanisms of action. This review summarizes the achievements in medicinal chemistry that enabled the translation of the ASO concept into real drugs, the molecular mechanisms of action of ASOs, the structure-activity relationship of ASO-protein binding, and the pharmacology, pharmacokinetics, and toxicology of ASOs. In addition, it discusses recent advances in medicinal chemistry in improving the therapeutic potential of ASOs by reducing their toxicity and enhancing their cellular uptake.