• Title/Summary/Keyword: small RNA (sRNA)

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Dual-Target Gene Silencing by Using Long, Synthetic siRNA Duplexes without Triggering Antiviral Responses

  • Chang, Chan Il;Kang, Hye Suk;Ban, Changill;Kim, Soyoun;Lee, Dong-ki
    • Molecules and Cells
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    • v.27 no.6
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    • pp.689-695
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    • 2009
  • Chemically synthesized small interfering RNAs (siRNAs) can specifically knock-down expression of target genes via RNA interference (RNAi) pathway. To date, the length of synthetic siRNA duplex has been strictly maintained less than 30 bp, because an early study suggested that double-stranded RNAs (dsRNAs) longer than 30 bp could not trigger specific gene silencing due to the induction of non-specific antiviral interferon responses. Contrary to the current belief, here we show that synthetic dsRNA as long as 38 bp can result in specific target gene silencing without non-specific antiviral responses. Using this longer duplex structure, we have generated dsRNAs, which can simultaneously knock-down expression of two target genes (termed as dual-target siRNAs or dsiRNAs). Our results thus demonstrate the structural flexibility of gene silencing siRNAs, and provide a starting point to construct multifunctional RNA structures. The dsiRNAs could be utilized to develop a novel therapeutic gene silencing strategy against diseases with multiple gene alternations such as viral infection and cancer.

Exosomes derived from microRNA-584 transfected mesenchymal stem cells: novel alternative therapeutic vehicles for cancer therapy

  • Kim, Ran;Lee, Seokyeon;Lee, Jihyun;Kim, Minji;Kim, Won Jung;Lee, Hee Won;Lee, Min Young;Kim, Jongmin;Chang, Woochul
    • BMB Reports
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    • v.51 no.8
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    • pp.406-411
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    • 2018
  • Exosomes are small membranous vesicles which contain abundant RNA molecules, and are transferred from releasing cells to uptaking cells. MicroRNA (miRNA) is one of the transferred molecules affecting the adopted cells, including glioma cells. We hypothesized that mesenchymal stem cells (MSCs) can secrete exosomes loading miRNA and have important effects on the progress of gliomas. To determine these effects by treating exosomal miRNA in culture media of miRNA mimic transfected MSCs, we assessed the in vitro cell proliferation and invasion capabilities, and the expression level of relative proteins associated with cell apoptosis, growth and migration. For animal studies, the mice injected with U87 cells were exposed to exosomes derived from miRNA-584-5p transfected MSCs, to confirm the influence of exosomal miRNA on the progress of glioma. Based on our results, we propose a new targeted cancer therapy wherein exosomes derived from miRNA transfected MSCs could be used to modulate tumor progress as the anticancer vehicles.

Effect of Corticosterone Administration on Small Intestinal Weight and Expression of Small Intestinal Nutrient Transporter mRNA of Broiler Chickens

  • Hu, X.F.;Guo, Yuming;Huang, B.Y.;Zhang, L.B.;Bun, S.;Liu, D.;Long, F.Y.;Li, J.H.;Yang, X.;Jiao, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.23 no.2
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    • pp.175-181
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    • 2010
  • The effects of corticosterone (CORT) administration on the weight of small intestine and the expression of nutrient transporter mRNA in the small intestine of broiler chickens (Gallus gallus domesticus) were investigated. One hundred and eight sevenday-old birds were randomly divided into two equal groups comprising a control group (CTRL) and an experimental group (CORT). CTRL birds were fed a basal diet and the CORT birds were fed a basal diet containing 30 mg corticosterone/kg from d 8 to 21. At 21 d of age, average daily feed intake (ADFI), serum corticosterone level, small intestinal absolute wet weight and relative weight, and relative abundance of SGLT1, CaBP-D28k, PepT1 mRNA in the duodenum and L-FABP mRNA in the jejunum were determined. The results showed that serum corticosterone level, liver weight and small intestinal relative weight (small intestinal wet weight/body weight) of CORT chickens were about 30.15%, 26.72% and 42.20% higher, respectively, than in the CTRL group (p<0.05). CORT birds had relative mRNA abundance of CaBP-D28k and PepT1 in the duodenum, and L-FABP in the jejunum which was 1.77, 1.37 and 1.94 fold higher, respectively, than in the CTRL group (p<0.05); the relative abundance of SGLT1 was 1.67 fold higher than in the CTRL group (p = 0.097). ADFI, small intestinal wet weight and length in CORT-treated broiler chickens was about 29.11%, 31.12% and 12.35% lower, respectively, than in the CTRL group (p<0.05). In conclusion, corticosterone administration lowered the wet weight but increased the relative weight of the small intestine and the expression of intestinal nutrient transporter mRNA of broiler chickens.

Using Reverse Dot Hybridization Method and 16S rRNA Gene (16S rDNA) for Identifying the Food Poisoning Microorganism in Foods (Reverse dot hybridization 방법과 16S rRNA gene(16S rDNA)을 이용한 식품에서 식중독균의 탐색)

  • Kim, Min-Seong;Shin, Kyu-Chul;Lee, Hyung-Gu;Han, Myung-Soo;Min, Byung-Re;Choi, Yong-Keel
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.470-474
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    • 2003
  • DNA sequence information on small-subunit rRNA gene (16S rDNA) obtained from food-poisoning bacterial culture was used to investigate the presence of bacterial pathogens in food. By reverse dot blot detection method, presence of food-poisoning bacteria could be confirmed on hybridization of digoxigenin-labeled 16S rDNA Polymerase Chain Reaction (PCR) primer product and biotin-labeled specific oligonucleotide probe. Escherichia coli, Bacillus cereus. and Salmonella sp. were used as the representative food-poisoning bacterial microorganisms. An oligonucleotide probe, based on the variable region of 16S rRNA gene, was used as the specific probe. These tools may be more useful than classic biochemical method for rapid identification of contaminated food.

Effects of antibacterial mouth rinses on multiple oral biofilms model (구강세정제가 다중 구강 바이오필름 모델에 미치는 영향)

  • Soo-Kyung Jun;Young-Suk Choi
    • Journal of Korean society of Dental Hygiene
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    • v.23 no.4
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    • pp.209-218
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    • 2023
  • Objectives: To confirm the antibacterial effects of each mouth rinse on multiple oral biofilms in vitro. Methods: The antibacterial effects of different mouth rinses were examined by ATP and counted colony forming units (CFU). Preformed oral biofilms on saliva coated hydroxyapatite (sHA) disks were treated with essential oil and saline; then, the multiple oral biofilms were observed by Scanning electron microscope (SEM). RNA sequencing analysis was performed on total RNA isolated from old biofilms of P. intermedia ATCC 49046. Results: In the CFU measured result compared to controls, preformed multiple oral biofilms were reduced from a low of 39.0% to 95.7% (p<0.05). The size of bacterial cells changed after treatment with the essential oil, and some of the cells ruptured into small pieces of cell debris. Gene expression in P. intermedia ATCC 49046 significantly altered in RNA transcribed and protein translated genes after exposure to essential oil. Conclusions: Mouth rinse solutions with different ingredients had different antibacterial effects and may alter surface structure and gene expression as determined by RNA sequencing.

Studies on Physicochemical and Biological Properties of Depolymerized Alginate from Sea tangle, Laminaria japonicus by Thermal Decomposition 5. Effects of Depolymerized Alginate on Body Weight, Organ, Pancreatic and Small Intestinal Composition, and Small Intestinal Microvilli Structure in Rats (다시마 (Laminaria japonicus) Alginate의 가열가수분해에 따른 물리$\cdot$화학적 및 생물학적 특성에 관한 연구 5. 랫드의 체중, 장기, 췌장과 소장의 성분 및 소장융모의 미세구조에 미치는 저분자 Alginate의 영향)

  • KIM Yuck-Yong;CHO Young-Je
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.34 no.1
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    • pp.21-26
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    • 2001
  • To examine functionality of depolymerized alginate obtained by hydrolysis of alginate through a heating process at $121^{\circ}C$ on gastrointestinal physiology, the changes of body weight, organ weight and length, pancreatic and small intestinal composition, and light microscopy (LM) observation of small intestinal microvilli's appearances were checked in the rats. Rats were fed diets containing $1\%, 5\%, and 10\%$ of each depolymerized alginate (HAG-10, HAG-50, HAG-100) and alginate for 35 days, The feeding of 5 and $10\%$ HAG-50 and $10\%$ alginate diets for 35 days significantly depressed the body weight gain, but increased the length and weight of the small intestine and cecum in rats (p<0.01). Pancreatic protease activity was decreased significantly (p<0.01) in all groups except lo/o of HAG-10 diets, but the protein content increased in all groups, However, pancreatic amylase and lipase activities as well as DNA and RNA content were not significantly different. The small intestinal protein and the DNA content were the highest in diets fed $5\%$ HAG-50; RNA content increased significantly (p<0.01) in all groups except in the fiber-free diets. Light microscopy (LM) observation showed growth of small intestinal microvilli with numerous ridges; the multiplication of the convolution goblet cells in rats fed with diets containing $5\%$ of HAG-50 were more than others group.

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Incredible RNA: Dual Functions of Coding and Noncoding

  • Nam, Jin-Wu;Choi, Seo-Won;You, Bo-Hyun
    • Molecules and Cells
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    • v.39 no.5
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    • pp.367-374
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    • 2016
  • Since the RNA world hypothesis was proposed, a large number of regulatory noncoding RNAs (ncRNAs) have been identified in many species, ranging from microorganisms to mammals. During the characterization of these newly discovered RNAs, RNAs having both coding and noncoding functions were discovered, and these were considered bifunctional RNAs. The recent use of computational and high-throughput experimental approaches has revealed increasing evidence of various sources of bifunctional RNAs, such as protein-coding mRNAs with a noncoding isoform and long ncRNAs bearing a small open reading frame. Therefore, the genomic diversity of Janusfaced RNA molecules that have dual characteristics of coding and noncoding indicates that the functional roles of RNAs have to be revisited in cells on a genome-wide scale. Such studies would allow us to further understand the complex gene-regulatory network in cells. In this review, we discuss three major genomic sources of bifunctional RNAs and present a handful of examples of bifunctional RNA along with their functional roles.

PCR and Restriction Fragment Pattern of 16S rRNA gene of Vibrio vulnificus (Vibrio vulnificus ATCC 27562의 16S rRNA 유전자의 PCR과 제한효소절단 방식)

  • 허문수;정초록
    • Journal of Life Science
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    • v.8 no.2
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    • pp.126-130
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    • 1998
  • A pair of designed primers (sequences from Gene Bank) amplified 16S fRNA gene of V. vulnificus within polymerase chain reaction (PCR) machine. This PCR product is about 1.3kb DNA fragment. Six enzymes (BamH I, Alu I, Sau3A I, Hind III, Sal I, Sma I) were used for restriction pattern analysis of amplified 16S rRNA gene of V. vulnificus ATCC 27562. Digested fragments are resolved by 3% agarose gel. BamH I did not show digested fragment so, there was no cutting site of BamH I in PCR product. Alu I produced three small fragments from 400 bp to 200 bp. Sau3A I produced three fragments larger than Alu I from 70 bp and 500 bp. One of fragments of Sal I was same with 500 bp of Hind III fragment and the other was 750 bp. Sma I showed two fragments of 800 bp and 470 bp. The profile of digested fragments of 16S rRNA of V.vulnificus ATCC 27562 will may be able to use standard profile for identification of V. vulnificus.

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Finding and Characterization of Viral Nonstructural Small Protein in Prospect Hill Virus Infected Cell

  • Nam, Ki-Yean;Chung, Dong-Hoon;Choi, Je-Won;Lee, Yun-Seong;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.221-233
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    • 1999
  • Prospect Hill Virus (PHV) is the well known serotype of hantavirus, a newly established genus in family Bunyaviridae. Extensive studies have upheld the original view of PHV genetics with three genes such as nucleocapsid (N) protein, envelope proteins (G1, G2) and RNA dependent RNA polymerase. In this study, we report the existence of additional gene that is encoded in an overlapping reading frame of the N protein gene within S genome segment of PHV. This gene is expected to encode a nonstructural small (NSs) protein and it seems to be only found in PHV infected cell. The presence and synthesis of NSs protein could be demonstrated in the cell infected with PHV using anti-peptide sera specific to the predicted amino acid sequence deduced from the second open reading frame. Ribosomal synthesis of this protein appears to occur at AUG codon at the 83rd base of S genome segment, downstream of N protein initiation codon. This protein is small in size (10.4 KDa) and highly basic in nature. The expression strategy of NSs protein appears that a signal mRNA is used to translate both N and NSs protein in PHV infected cell. 10 KDa protein in virus infected cell lysates can bind to mimic dsRNA. This fact strongly suggests that NSs protein may be involved in virus replication on late phase of viral life cycle.

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Characterization of a Tomato (Lycopersicon esculentum Mill.) Ripening-associated Membrane Protein (TRAMP) Gene Expression and Flavour Volatile Changes in TRAMP Transgenic Plants

  • Kim Seog-Hyung;Ji Hee-Chung;Lim Ki-Byung
    • Journal of Plant Biotechnology
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    • v.7 no.2
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    • pp.87-95
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    • 2005
  • The tomato ripening associated membrane protein (TRAMP) (Fray et al., 1994) is a member of the major intrinsic protein (MIP) family, defined as channels facilitating the passage of water and small solutes through membranes. During normal fruit ripening the TRAMP mRNA levels were increased whereas the expression levels of TRAMP in low ethylene ACO1-sense suppressed lines, Nr and rin fruits, were lower than at the breaker stage of wild type fruit. TRAMP mRNA is inhibited by $LaCl_3$, which is an inhibitor of $Ca^{2+}$-stimulated responses, treatment but drought condition did not affect TRAMP expression. The levels of TRAMP mRNA transcripts were substantially higher in the dark treated seedlings and fruits. These suggest that TRAMP function as a water channel may be doubted because of several reasons; no water content was changed during ripening in wild type, antisense and overexpression lines, TRAMP expression under light condition was lower than dark condition and TRAMP expression was not changed in drought condition. Co-suppression plant, 3588 was one of sense suppression lines, which contain CaMV 35S promoter and sense pNY507 cDNA, produced small antisense RNA, approximately 21-25 nucleotides in length, mediated post-transcriptional gene silencing. Therefore, TRAMP expression was inhibited by small antisense and multiple copies might induce gene silencing without any production of double strand RNA. Total seven selected volatile productions, isobutylthiazole, 6-methyl-5-hepten-2-one, hexanal, hexenal methylbutanal, hexenol, and methylbutanol, were highly reduced in sense line whereas total volatile production was increased in TRAMP antisense line. These results suggested TRAMP might change volatile related compounds.