• 대한의생명과학회지
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• 제29권4호
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• pp.231-241
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• 2023

Messenger RNA (mRNA)-based vaccines and treatments have recently emerged as a promising strategy. Naked mRNA presents various limitations for direct delivery. Therefore, in this paper, Lipid Nanoparticles (LNPs) were utilized for the delivery of mRNA. Lipid nanoparticle (LNP) mRNA systems are highly effective as vaccines, but their efficacy for pulmonary delivery has not yet been fully established. Additionally, research on effective delivery systems and administration methods for vaccines is required to resolve the stability and degradation issues associated with naked mRNA delivery. This study aimed to determine mRNA delivery efficiency via the inhalation of a lipid nanoparticle (LNP) formulation designed specifically for pulmonary delivery. To this purpose, we built a library of seven LNP configurations with different lipid molar and N/P ratios and evaluated their encapsulation efficiency using gel retardation assay. Among the tested LNPs, LNP1, LNP2-2, and LNP3-2 demonstrated high transfection efficiency in vitro based on FACS analyses luciferase assays, and intracellular accumulation tests. The mRNA delivery efficiencies of the selected LNPs after inhalation and intravenous injection were compared and evaluated. LNP2-2 showed the highest mRNA expression in healthy mouse lungs when aerosolized and was found to be non-toxic. These results indicate that LNP2-2 is a promising carrier for lung mRNA delivery via inhalation.

• 미생물학회지
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• 제26권1호
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• pp.27-31
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• 1988

Biosynthesis and processing of cytoplasmic mRNA from heterogenous nuclear RNA (hn-RNA) in Aspergillus phoenicis were studied by $^{3}H$-uridine labeling and synchronous culture techniques during their life cycle. Incorporations of $^{3}H$-uridine into hn-RNA and mRNA were most rapid in vesicle-phialide fromation stage and diminished in hyphal growth stage. The processing of cytoplasmic mRNA from hn-RNA was proceeded more rapidly in hyphal growth and conidiophore formation stages than in conidia and vesicle-phialide formation stages. The specific radioactivities of hn-RNA and mRNA were very high in vesicle-phialide formation stage.

• 한국임상수의학회지
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• 제40권2호
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• pp.164-174
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• 2023

Canine splenic tumors (STs) are commonly diagnosed during imaging examinations, such as in X-ray and ultrasonography examinations, suggesting their higher prevalence, especially in older dogs. Despite this high prevalence, there are no effective treatment options for STs because of the difficulties in determining therapeutic targets. However, recently, the importance of microRNAs (miRNAs) has evolved owing to their ambivalent characteristics. Biomarkers and novel therapies using miRNAs have been well-studied in human cancer research compared to canine research, except for mammary gland tumors. Therefore, this study aimed to comparatively analyze miRNA expression profiles according to malignancy and biopsy sites to identify novel therapeutic and diagnostic targets. Tissue samples were collected directly from splenic tumor masses and immersed in RNAlater solution for further analysis. To investigate differentially expressed genes (DEGs) between tumor and normal tissues, we used RNA-seq and miRNA microarray analysis. Then, functional analysis based on DEGs was conducted to sort tumor-related DEGs. We found that cfa-miR-150 was upregulated in benign tumors, whereas cfa-miR-134 was upregulated in malignant tumors. Despite limited information on canine miRNAs, we identified two potential biomarkers for the differential diagnosis of STs.

• Molecules and Cells
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• 제42권5호
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• pp.379-385
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• 2019

Non-coding RNAs (ncRNAs) comprise various RNA species, including small ncRNAs and long ncRNAs (lncRNAs). ncRNAs regulate various cellular processes, including transcription and translation of target messenger RNAs. Recent studies also indicate that ncRNAs affect organismal aging and conversely aging influences ncRNA levels. In this review, we discuss our current understanding of the roles of ncRNAs in aging and longevity, focusing on recent advances using the roundworm Caenorhabditis elegans. Expression of various ncRNAs, including microRNA (miRNA), tRNA-derived small RNA (tsRNA), ribosomal RNA (rRNA), PIWI-interacting RNA (piRNA), circular RNA (circRNA), and lncRNA, is altered during aging in C. elegans. Genetic modulation of specific ncRNAs affects longevity and aging rates by modulating established aging-regulating protein factors. Because many aging-regulating mechanisms in C. elegans are evolutionarily conserved, these studies will provide key information regarding how ncRNAs modulate aging and lifespan in complex organisms, including mammals.

• BMB Reports
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• 제50권11호
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• pp.554-559
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• 2017

MicroRNAs (miRNAs) are ~22nt-long single-stranded RNA molecules that form a RNA-induced silencing complex with Argonaute (AGO) protein to post-transcriptionally downregulate their target messenger RNAs (mRNAs). To understand the regulatory mechanisms of miRNA, discovering the underlying functional rules for how miRNAs recognize and repress their target mRNAs is of utmost importance. To determine functional miRNA targeting rules, previous studies extensively utilized various methods including high-throughput biochemical assays and bioinformatics analyses. However, targeting rules reported in one study often fail to be reproduced in other studies and therefore the general rules for functional miRNA targeting remain elusive. In this review, we evaluate previously-reported miRNA targeting rules and discuss the biological impact of the functional miRNAs on gene-regulatory networks as well as the future direction of miRNA targeting research.

• Molecules and Cells
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• 제44권7호
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• pp.538-539
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• 2021

• Molecules and Cells
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• 제39권12호
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• pp.841-846
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• 2016

Local protein synthesis mediates precise spatio-temporal regulation of gene expression for neuronal functions such as long-term plasticity, axon guidance and regeneration. To reveal the underlying mechanisms of local translation, it is crucial to understand mRNA transport, localization and translation in live neurons. Among various techniques for mRNA analysis, fluorescence microscopy has been widely used as the most direct method to study localization of mRNA. Live-cell imaging of single RNA molecules is particularly advantageous to dissect the highly heterogeneous and dynamic nature of messenger ribonucleoprotein (mRNP) complexes in neurons. Here, we review recent advances in the study of mRNA localization and translation in live neurons using novel techniques for single-RNA imaging.

• Molecules and Cells
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• 제37권1호
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• pp.1-8
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• 2014

Mammalian-cell messenger RNAs (mRNAs) are generated in the nucleus from precursor RNAs (pre-mRNAs, which often contain one or more introns) that are complexed with an array of incompletely inventoried proteins. During their biogenesis, pre-mRNAs and their derivative mRNAs are subject to extensive cis-modifications. These modifications promote the binding of distinct polypeptides that mediate a diverse array of functions needed for mRNA metabolism, including nuclear export, inspection by the nonsense-mediated mRNA decay (NMD) quality-control machinery, and synthesis of the encoded protein product. Ribonucleoprotein complex (RNP) remodeling through the loss and gain of protein constituents before and after pre-mRNA splicing, during mRNA export, and within the cytoplasm facilitates NMD, ensuring integrity of the transcriptome. Here we review the mRNP rearrangements that culminate in detection and elimination of faulty transcripts by mammalian-cell NMD.

• 한국동물학회지
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• 제33권2호
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• pp.183-190
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• 1990

본 연구는 prolactin(PRL)유전자 발현, 분비의 생리적 변화와 성주기 특정 시기의 PRL mRNA수준 및 분비에 미치는 내인성 오피오이드의 영향을 조사하였다. 최소한 두번의 연속적인 성주기를 거친 성숙한 흰쥐에서 성주기의 각 시기(10:00시)에, proestrus시기에는 10:00-20:00 시동안에는 2시간 간격으로 도살하였고, naloxone (2mg/kg b.w.)은 도살 30분전에 피하주사 하였다. PRL mRNA의 수준의 흰쥐의 PRL cDNA를 probe로 하여 RNA-blot hybridization방법에 의해서, 혈중 PRL농도 변화는 방사면역측정법에 의해 측정하였다. 뇌하수체 PRL mRNA의 수준과 혈중 PRL수준은 diestrus I, II and proestrus그리고 estrus시기의 10:00시에는 급격한 변화를 보이이 않았다. 이때 naloxone처리는 영향을 미치지 못했다. proestrus시기를 세분화하여 조사한 결과 PRL mRNA의 수준은 정오에 최고 수준에 도달하였고, 오후 6:00까지 점차적으로 감소하였다. 그후 8:00시에 다시 증가하였다. estrus동안 naloxone은 혈중의 PRL수준을 명백히 억제하였으나 PRL mRNA수준에는 영향이 없었다. proestrus시기 동안 혈중 PRL변화와 뇌하수체 PRL mRNA변화는 서로 상이하게 조절되며,PRL mRNA수준이 흰쥐 성주기 동안 변화하고 있는 사실에서 PRL 유전자 발현이 생리적으로 조절되고 있음을 시사한다.

• BMB Reports
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• 제55권3호
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• pp.125-135
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• 2022

Continuously renewing the proteome, translation is exquisitely controlled by a number of dedicated factors that interact with the ribosome. The RNA helicase DDX3 belonging to the DEAD box family has emerged as one of the critical regulators of translation, the failure of which is frequently observed in a wide range of proliferative, degenerative, and infectious diseases in humans. DDX3 unwinds double-stranded RNA molecules with coupled ATP hydrolysis and thereby remodels complex RNA structures present in various protein-coding and noncoding RNAs. By interacting with specific features on messenger RNAs (mRNAs) and 18S ribosomal RNA (rRNA), DDX3 facilitates translation, while repressing it under certain conditions. We review recent findings underlying these properties of DDX3 in diverse modes of translation, such as cap-dependent and cap-independent translation initiation, usage of upstream open reading frames, and stress-induced ribonucleoprotein granule formation. We further discuss how disease-associated DDX3 variants alter the translation landscape in the cell.