• Molecules and Cells
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• 제27권6호
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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.

• The Plant Pathology Journal
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• 제16권5호
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• pp.252-256
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• 2000

The 16S-23S rRNA intergenic spacer regions (ISRs) were sequenced and analyzed to design specific primer for identification of Pectobacterium chrysanthemi. Two types ISRs, large and small ISRs, were identified from three strains (ATCC 11663, KACC 10163 and KACC 10165) of P. chrysanthemi and Pectobacterium carotovorum subsp. carotovorum ATCC 15713.Large ISRs contained transfer RNA-Ile(tRNA$^{Ile}$)and tRNA$^{Ala}$, and small ISRs contained tRNA$^{Glu}$. Size of the small ISRs of P. chrysanthemi ranged on 354-356 bp, while it was 451 bp in small ISR of P. carotovorum subsp. carotovorum ATCC 15713. From hypervariable region of small ISRs, species-specific primer for P. chrysanthemi with 20 bp length (CHPG) was designed from hypervariable region of small ISRs, which was used as forward promer to detect P. chrysanthemi strains with R23-1R produced PCR product of about 260bp size (CHSF) only from P. chrysanthemi strains, not from other Pectobacterium spp. and Erwinia spp. Direct PCR from bacterial cell without extracting DNA successfully amplified a specific fragment, CHSF, from P. chrysanthemi ATCC 11663. The limit of PCR detection was 1${\pm}10^2$ cfu/ml.

• 미생물학회지
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• 제30권3호
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• pp.177-180
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• 1992

Among 120 U. maydis strains isolated in Korea 14 different strains containing specific viral dsRNA segments were analyzed for the distribution of dsRNA and the production of toxin protein. Several distinctive dsRNA patterns were identified, 9 cases of P type with typical H, M and L ds RNA and one case of non-P-type, the frequency of a specific isolate was decreased with increasing number of dsRNA segments. The presence of dsRNA had no effect on the cultural or morphological phenotype of the host. Two isolates containing P type dsRNA segments appeared to produce toxin protein (killer strains) which inhibited the growth of 4 isolates (sensitive strain) with different susceptibility. Two killer strains contain unique M dsRNA segment which may code for toxin protein. However, the presence of toxin-sensitive strains among dsRNA-free isolates was similar to that of ds RNA containing strains.

• 한국동물학회지
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• 제20권1호
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• pp.9-18
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• 1977

RNA 분해효소에 저항하는 RNA 분자들이 양서류의 난에 존재하는지의 여부를 조사하기 전에 필요한 몇가지 예비실험을 하기 위하여 Xenopus laevis에 난에서 RNA를 추출 하였다. Sephadex G-100 column chromatography는 세 개의 peak을 항상 보여주고 있다. 첫째 peak에 포함되어있는 고분자량의 RNA만을 $^{3}H$-dimethyl sulfate를 사용하여 시험관내에서 label하여 tRNA로부터의 base paired oligonucleotide의 참여를 배제하였다. 이 방법으로 아주 높은 specific activity를 얻을 수 있었으며 또한 부착된 methyl group은 대단히 안정성을 보였다.

• 대한화학회지
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• 제65권2호
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• pp.121-124
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• 2021

RNA molecules which bind to the G-rich sequence in the 5'-UTR RNA which plays an important role in expression of N-ras, were selected. The secondary structures of five selected RNA aptamers including primer sequence were found by the CLC RNA workbench ver. 4.2 program (www.clcbio.com) and investigated with RNA structural probes such as RNase T1 which has specificity for a G in single-stranded region, RNase V1 specific for double strand and nuclease S1 specific for single strand. The generalized secondary structure model was proposed and characterized. It was composed of a central long double strand region flanked by single strand region at both end sides. The double strand region had an internal single-strand region and bulges. The single strand loop in the right side was composed of four or five nucleotides.

• 한국응용곤충학회지
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• 제56권2호
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• pp.153-164
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• 2017

이중나선형 RNA (dsRNA)를 이용한 유전자 발현 억제기술이 다양한 생명체에서 기능 유전체학을 연구하는 데 이용되고 있다. 이 기술의 원리는 전사후 단계에서 유전자 발현을 조절하는 RNA 간섭에 기인된다. dsRNA를 이용하여 특정 유전자의 발현 억제는 심각한 치사효과를 줄 수 있다. 이러한 분자기작을 해충 방제에 적용하여 특정 dsRNA를 이용한 새로운 살충제를 개발하고 있다. 본 종설은 dsRNA를 이용한 RNA간섭 원리를 설명하고 이를 이용한 해충 방제를 구현한 여러 예를 살펴본다. 그리고 해충방제를 실현시키기 위해 현재 이 기술이 담고 있는 한계를 고찰하고 이에 대한 대응 방안을 본 종설에서 제공하고자 한다.

• Bioinformatics and Biosystems
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• 제2권1호
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• pp.17-23
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• 2007

Short interfering RNA(siRNA)는 특별한 gene의 발현을 막는데 사용될 수 있고 그 gene의 기능과 치료의 적용에 많은 가능성을 가지고 있지만, 효과적인 siRNA를 디자인하는 방법은 아직까지 명확하지 않다. 효과적인 siRNA는 서열적인 경향을 가지고 있는데 낮은 G/C content, Sense strand의 3' 끝에 적은 안정성과 1번 위치에는 G/C, 19번 위치에는 A/U의 존재 여부를 들 수 있다. 이러한 특성 말고도 최근에는 mRNA의 2차구조가 RNAi 작용에 중요한 역할을 하게 되는데 복잡한 구조(hairpin, multi loop)를 가지고 수소결합을 많이 하여 안정한 상태에 있는 부분은 siRNA의 기능을 크게 줄어들게 한다. 또한, siRNA가 특정한 mRNA에 작동하도록 BLAST 검색을 하여 부작용의 가능성을 배제한다.

• Journal of Microbiology and Biotechnology
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• 제28권3호
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• pp.367-374
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• 2018

RNA interference provides an effective tool for developing antitumor therapies. Cell-penetrating peptides (CPPs) are delivery vectors widely used to efficiently transport small-interfering RNA (siRNA) to intracellular targets. In this study, we investigated the efficacy of the cancer-specific CPP carrier BR2 to specifically transport siRNA to cancer-target cells. Our results showed that BR2 formed a complex with anti-vascular endothelial growth factor siRNA (siVEGF) that exhibited the appropriate size and surface charge for in vivo treatment. Additionally, the BR2-VEGF siRNA complex exhibited significant serum stability and high levels of gene-silencing effects in vitro. Moreover, the transfection efficiency of the complex into a cancer cell line was higher than that observed in non-cancer cell lines, resulting in downregulated intracellular VEGF levels in HeLa cells and comprehensively improved antitumor efficacy in the absence of significant toxicity. These results indicated that BR2 has significant potential for the safe, efficient, and specific delivery of siRNA for diverse applications.

• Genomics & Informatics
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• 제16권4호
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• pp.28.1-28.6
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• 2018

Due to the increasing interest in synonymous codons, several codon bias-related terms were introduced. As one measure of them, the tRNA adaptation index (tAI) was invented about a decade ago. The tAI is a measure of translational efficiency for a gene and is calculated based on the abundance of intracellular tRNA and the binding strength between a codon and a tRNA. The index has been widely used in various fields of molecular evolution, genetics, and pharmacology. Afterwards, an improved version of the index, named specific tRNA adaptation index (stAI), was developed by adapting tRNA copy numbers in species. Although a subsequently developed webserver (stAIcalc) provided tools that calculated stAI values, it was not available to access pre-calculated values. In addition to about 100 species in stAIcalc, we calculated stAI values for whole coding sequences in 148 species. To enable easy access to this index, we constructed a novel web database, named STADIUM (Species-specific tRNA adaptive index compendium). STADIUM provides not only the stAI value of each gene but also statistics based on pathway-based classification. The database is expected to help researchers who have interests in codon optimality and the role of synonymous codons. STADIUM is freely available at http://stadium.pmrc.re.kr.

• BMB Reports
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• 제48권3호
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• pp.147-152
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• 2015

Small interfering RNA (siRNA) functions through pairing with specific mRNA sequences and results in the mRNA's degradation. It is a potential therapeutic approach for many diseases caused by altered gene expression. The delivery of siRNA is still a major problem due to its rapid degradation in the circulation. Various strategies have been proposed to help with the cellular uptake of siRNA and short or small hairpin RNA (shRNA). Here, we reviewed recently published data regarding local applications of siRNA. Compared with systemic delivery methods, local delivery of siRNA/shRNA has many advantages, such as targeting the specific tissues or organs, mimicking a gene knockout effect, or developing certain diseases models. The eye, brain, and tumor tissues are 'hot' target tissues/organs for local siRNA delivery. The siRNA can be delivered locally, in naked form, with chemical modifications, or in formulations with viral or non-viral vectors, such as liposomes and nanoparticles. This review provides a comprehensive overview of RNAi local administration and potential future applications in clinical treatment.