• Title/Summary/Keyword: Gene delivery system

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Research Trends in Bifidobacterium (비피더스 연구동향)

  • Park, Myeong-Soo;Ji, Geun-Eog
    • KSBB Journal
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    • v.25 no.4
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    • pp.319-329
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    • 2010
  • Bifidobacteria comprises up to 25% of the cultivable fecal bacteria in adults and 80% in infants. Many in vivo and clinical research results supporting its efficacy in the prevention and improvement of gastrointestinal health have been accumulated. As a consequence, expert committee WHO/FAO expert committee recommended Bifidobacterium as representative probiotics together with Lactobacillus acidophilus. In this review, research trends in bifidobacteria concerning the classification and identification of the genus Bifidobacterium, modulation of intestinal microflora, improvement of constipation, prevention of diarrhea, alleviation of atopy and allergy, barrier function through antimicrobial activity andimmune enhancement of the host will be introduced. Several gene expression systems based on bifidobacterial plasmids have been developed and successfully used to express several heterologous genes including anticancer proteins in Bifidogacterium. In animal test, bifidobacteria was proven to be a promising candidate for safe gene delivery system which can specifically colonize in the solid tumor.

Photo-controlled gene expression by fluorescein-labeled antisense oligonucleotides in combination with visible light irradiation

  • Ito, Atsushi;Kaneko, Tadashi;Miyamoto, Yuka;Ishii, Keiichiro;Fujita, Hitoshi;Hayashi, Tomonori;Sasaki, Masako
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.451-453
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    • 2002
  • A new concept of "photo" -antisense method has been evaluated, where the inhibition of gene expression by the conventional antisense method is enhanced by photochemical binding between antisense oligonucleotides conjugated with photo-reactive compound and target mRNA or DNA. Fluorescein labeled oligodeoxyribonucleotides (F-DNA) was delivered to cell nuclei in the encapsulated form in multilamellar lecithin liposomes with neutral charge. F-DNA was previously shown to photo-bind to the complementary stranded DNA, and the delivery system using neutral liposome to be effective in normal human keratinocytes. In the present study, we used human kidney cancer G401.2/6TG.1 cell line to be advantageous in reproducible experiments. p53 was adopted as a target gene since antisense sequence information has been accumulated. The nuclear localization ofF-DNA was identified by comparing the fluorescence ofF-DNA with that of Hoechst 33258 under fluorescence microscope. After 7hr incubation to accumulate p53 protein induced by UV -B, p53 protein was quantified by Western blot. After 2hrs from F-DNA application, about 30% of cell population incorporated F-DNA in their nuclei with some morphological change possibly due to liposomal toxicity. Irradiation of visible light longer than 400nm from solar simulator at this time enhanced the inhibitory action of antisense F-DNA. The present results suggest that photo-antisense method is promising to control gene expression in time and space dependent manner. Further improvement of F-DNA delivery to cancer cells in the stability and toxicity is in progress. progress.

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Systemic Analysis of a Novel Coxsackievirus Gene Delivery System in a Mouse Model

  • Kim, Yeon-Jung;Yun, Soo-Hyeon;Lim, Byung-Kwan;Park, Ki-Bum;Na, Ha-Na;Jeong, Soo-Young;Kim, Dae-Sun;Cho, Young-Joo;Jeon, Eun-Seok;Nam, Jae-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.307-313
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    • 2009
  • In order to systemically investigate the possibility of using coxsackievirus B3 (CVB3) to deliver foreign genes in vivo, a recombinant strain of CVB3 encoding the renilla gene (CVB3-renilla) was constructed. The recombinant CVB3 resulted in extensive and transient expression of the renilla protein within mouse organs, especially the pancreas. The level of expression was generally dependent upon the viral titer present. Moreover, the CVB3-renilla strain was completely attenuated. Interestingly, the recombinant CVB3 vector was expressed much more strongly in mouse organs than was a comparable adenoviral vector. The CVB3-renilla strain did not express the renilla gene in mice with pre-existing coxsackievirus-specific neutralizing antibodies, but direct organ-specific administration of the virus during open-peritoneum surgery was able to circumvent this immunity. This coxsackievirus vector may represent a useful means for delivering and expressing foreign genes in mouse models in an acute and extensive fashion.

Establishment of a Simple and Rapid Gene Delivery System for Cucurbits by Using Engineered Zucchini Yellow Mosaic Virus

  • Kang, Minji;Seo, Jang Kyun;Choi, Hoseong;Choi, Hong Soo;Kim, Kook Hyung
    • The Plant Pathology Journal
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    • v.32 no.1
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    • pp.70-76
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    • 2016
  • The infectious full-length cDNA clone of zucchini yellow mosaic virus (ZYMV) isolate PA (pZYMV-PA), which was isolated from pumpkin, was constructed by utilizing viral transcription and processing signals to produce infectious in vivo transcripts. Simple rub-inoculation of plasmid DNAs of pZYMV-PA was successful to cause infection of zucchini plants (Cucurbita pepo L.). We further engineered this infectious cDNA clone of ZYMV as a viral vector for systemic expression of heterologous proteins in cucurbits. We successfully expressed two reporter genes including gfp and bar in zucchini plants by simple rub-inoculation of plasmid DNAs of the ZYMV-based expression constructs. Our method of the ZYMV-based viral vector in association with the simple rub-inoculation provides an easy and rapid approach for introduction and evaluation of heterologous genes in cucurbits.

Molecular Application in Psychiatry (정신과의 분자생물학 적용)

  • Choi, Ihn-Geun
    • Korean Journal of Biological Psychiatry
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    • v.7 no.2
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    • pp.115-122
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    • 2000
  • The development of molecular biology has brought many changes in psychiatry. Molecular biology makes us possible to know the cause of mental disorders that provide the way to prevent the disorders, and to develop various accurate diagnostic and treatment methods for mental disorders. The author discusses the concept, cause, and treatment of mental disorders in the aspect of molecular biology. Importing the methods of molecular biology into psychiatry, we can anticipate to get a number of the goals of psychiatric genetics, including identification of specific susceptibility genes, clarification of the pathophysiological processes whereby these genes lead to symptoms, establishment of epigenetic factors that interact with these genes to produce disease, validation of nosological boundaries that more closely reflect the actions of these genes, and development of effective preventive and therapeutic interventions based on genetic counseling, gene therapy, and modification of permissive or protective environmental influences. In addition to their capacity to accelerate the discovery of new molecules participating in the nervous system's response to disease or to self-administered drugs, molecular biological strategies can also be used to determine how critical a particular gene product may be in mediating a cellular event with behavioral importance. Molecular biology probably enables us discover the environmental factors of mental disorders and allow rational drug design and gene therapies for mental disorders, by isolation of gene products that facilitate a basic understanding of the pathogenesis of these disorders. A specific genetic linkage may suggest a novel class of drugs that has not yet been tried. With respect to gene therapy, the hypothetical method would use a gene delivery system, most likely a modified virus, to insert a functional copy of a mutant gene into those brain cells that require the gene for normal function.

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Applications of Transposon-Based Gene Delivery System in Bacteria

  • Choi, Kyoung-Hee;Kim, Kang-Ju
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.217-228
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    • 2009
  • Mobile genetic segments, or transposons, are also referred to as jumping genes as they can shift from one position in the genome to another, thus inducing a chromosomal mutation. According to the target site-specificity of the transposon during a transposition event, the result is either the insertion of a gene of interest at a specific chromosomal site, or the creation of knockout mutants. The former situation includes the integration of conjugative transposons via site-specific recombination, several transposons preferring a target site of a conserved AT-rich sequence, and Tn7 being site-specifically inserted at attTn7, the downstream of the essential glmS gene. The latter situation is exploited for random mutagenesis in many prokaryotes, including IS (insertion sequence) elements, mariner, Mu, Tn3 derivatives (Tn4430 and Tn917), Tn5, modified Tn7, Tn10, Tn552, and Ty1, enabling a variety of genetic manipulations. Randomly inserted transposons have been previously employed for a variety of applications such as genetic footprinting, gene transcriptional and translational fusion, signature-tagged mutagenesis (STM), DNA or cDNA sequencing, transposon site hybridization (TraSH), and scanning linker mutagenesis (SLM). Therefore, transposon-mediated genetic engineering is a valuable discipline for the study of bacterial physiology and pathogenesis in living hosts.

Preparation and Characterization of O-Carboxymethyl Chitosan Ion-complexed Poly(L-Lysine) for Drug and Gene Delivery System (약물 및 유전자 전달체로 응용하기 위한 Poly(L-Lysine)이 결합된 O-Carboxymethyl Chitosan PEG의 제조와 특성)

  • Nam, Joung-Pyo;Kim, Young-Min;Park, Jin-Su;Lee, Eung-Jae;Choi, Chang-Yong;Jang, Mi-Kyeong;Nah, Jae-Woon
    • Applied Chemistry for Engineering
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    • v.21 no.6
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    • pp.643-647
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    • 2010
  • O-carboxymethyl water-soluble chitosan (OCMCh) prepared for enhance the application of chitosan was modified with mthoxy polyethyleneglycol (mPEG) by ion-complex for long circulation in the blood. OCMCh-PEG-PLLs was prepared by forming ion-complex with OCMCh-PEG and Poly(L-Lysine) (PLL) for drug and gene delivery system. The physicochemcal characterisitcs of OCMCh-PEG-PLLs were investigated by FT-IR, $^1H$-NMR. These results showed that CMCh-PEG-PLLs were successfully syntehsized by ion-complex. Particle size distribution and zeta potential of the OCMCh-PEG-PLLs were determined using dynamic light scattering technique. Transmission electron microscopy (TEM) was also used to observe the morphology of the OCMCh-PEG-PLLs. OCMCh-PEG-PLLs have spherical shapes with particle size 290∼390 nm. OCMCh-PEG-PLLs were showed when the feeding amount of mPEG ratio was increased, particle size and zeta potential were decreased. Based on these results, it is possible to introduction of the OCMCh-PEG-PLLs into various biomedical fields such as drug and gene delivery system.

Genetic and Epigenetic Biomarkers on the Personalized Nutrition

  • An Sung-Whan
    • Proceedings of the Korean Society of Food Science and Nutrition Conference
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    • 2004.11a
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    • pp.271-274
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    • 2004
  • Nutritional genomics is a new field of study of how nutrition interacts with an individual's genome or individual responds to individual diets. Systematic approach of nutritional genomics will likely provide important clues about responders and non-responders. The current interest in personalizing health stems from the breakthroughs emerging in integrative technologies of genomics and epigenomics and the identification of genetic and epigentic diversity in individual's genetic make-up that are associated with variations in many aspects of health, including diet-related diseases. Microarray is a powerful screen system that is being also currently employed in nutritional research. Monitoring of gene expression at genome level is now possible with this technology, which allows the simultaneous assessment of the transcription of tens of thousands of genes and of their relative expression of pathological cells such tumor cells compared with that of normal cells. Epigenetic events such as DNA methylation can result in change of gene expression without involving changes in gene sequence. Recent developed technology of DNAarray-based methylation assay will facilitate wide study of epigenetic process in nutrigenomics. Some of the areas that would benefitfrom these technologies include identifying molecular targets (Biomarkers) for the risk and benefit assessment. These characterized biomarkers can reflect expose, response, and susceptibility to foods and their components. Furthermore the identified new biomarker perhaps can be utilized as a indicator of delivery system fur optimizing health.

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The Synthesis of Artery Wall Targeted Gene Carrier Using Low Molecular Water-Soluble Chitosan (저분자량 수용성 키토산을 이용한 동맥 벽 표적성 유전자 전달체의 합성)

  • Choi Chang-Yong;Jang Mi-Kyeong;Nah Jae-Woon
    • Polymer(Korea)
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    • v.30 no.4
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    • pp.279-285
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    • 2006
  • Non-viral gene carriers continue to attract a great deal of interest due to advantageous safety profile. Among the non-viral gene carriers, cationic liposomes or synthetic gene carriers are efficient DNA carriers in vitro. but their in vivo applications are greatly hampered because of low biocompatibility. On the other hand, chitosan, a natural cationic polysaccharide, is a candidate non-viral vector for gene delivery because of its low cytotoxicity and high positive charges. In this work, targeted gene carrier was synthesized to target artery wall cells using low molecular water-soluble chitosan (LMWSC). The molecular weight $(M_W)$ and degree of de acetylation (DDA) of LMWSC were measured by relative viscometer and Kina titration. respectively. The structure of LMWSC was analyzed by measuring FTIR, $^1H-NMR,\;and\;^{13}C-NMR$. AWBP-PEG-g-LMWSC was synthesized by conjugation of the artery wall binding peptide (AWBP), a specific targeting peptide, to the end of pegylated LMWSC as a gene carrier to target artery wall cells. The synthesized AWBP-PEG-g-LMWSC were analyzed by measuring FTIR, $^1H-NMR$, zeta -potentiometer, and atomic force microscopy (AFM).

In vitro Nasal Cell Culture Systems for Drug Transport Studies

  • Cho, Hyun-Jong;Termsarasab, Ubonvan;Kim, Jung-Sun;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.321-332
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    • 2010
  • Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the "passage" culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.