• Korean Journal of Agricultural Science
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• v.34 no.1
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• pp.47-52
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• 2007

유전자의 발현이 다양한 형태로 억제되는 것을 silencing이라고 한다. 유전자 발현억제 방법 중 antisense RNA는 자연 상태의 mRNA에 역상보적인 RNA 분자로서 형질전환된 세포에서 그 mRNA의 전이을 억제하는 데 이용된다. RNA분자에 대하여 상보적 염기배열을 갖는 RNA는 분자간 결합을 연결하여 RNA의 기능 발현에 억제적으로 작용한다고 생각된다. 미생물에 나타나는 유전자발현 제어기작으로서 어느 특정한 mRNA에 대하여 상보적인 RNA가 유전자 발현의 억제인자로서 작용하고 있는 예가 몇 가지 알려져 있다. 이러한 경우 antisense RNA는 mRNA 상의 전이개시영역과 상보적 배열을 하고 있고, 전이과정을 방해한다고 추정되고 있지만 작용기작의 상세한 내용은 아직 명확하지가 않다. 한편 안티센스RNA는 임의의 표적유전자에 대하여 인위적으로 제작할 수가 있기 때문에 인위적인 유전자발현제어의 한 방법으로 이용되고 있다. 특정한 유전자에 대한 antisense RNA를, 발현하는 유전자를 인위적으로 제작하여 세포내에 도입하면 표적유전자의 발현을 특이적으로 억제 제어할 수 있는 것이 기대되어, 다양한 생물체를 대상으로 하여 많은 시도가 이루어지고 있으며 몇 가지 성공적인 보고가 있다. 그 중 식물이차대사과정에 관련 유전자를 대상으로 antisense RNA 기법으로 유전자의 발현억제와 이차대사산물 생산조절에 관한 연구를 이 논문에서 조사하고 정리하였다.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.3
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• pp.337-344
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• 2018

Hepatocellular carcinoma (HCC), a major type of hepatoma, is associated with high recurrence and mortality because of its uncontrolled metastatic feature. Silymarin is a polyphenolic flavonoid from Silybum marianun (milk thistle) and exhibits anti-carcinogenic activity through modulation of the epithelial-mesenchymal transition (EMT) in several cancer cells. In this study, the inhibitory mechanism of silymarin against migration and invasion was investigated in the Huh7 HCC cell line. Wound healing and in vitro invasion assays were conducted to examine the effects of silymarin on migration and invasion. Western blot analysis was also applied to evaluate the inhibitory effects of silymarin on the EMT-related genes and their upstream signaling molecules. Silymarin inhibited the migratory and invasive activities of Huh7 cells. In addition, silymarin attenuated the protein expression levels of vimentin and matrix metalloproteinase (MMP)-9 as well as their transcription factors, Snail, and nuclear factor $(NF)-{\kappa}B$, while the expression of E-cadherin was increased by the silymarin treatment. Among the upstream signaling molecules, the phosphorylation of Akt was inhibited by the silymarin treatment, which was confirmed by the selective inhibitor, LY294002. Consequently, silymarin inhibited the invasive and migratory activities in Huh7 cells through the modulation of EMT-related gene expression by the PI3K/Akt signaling pathway, which may have potential as a chemopreventive agent against HCC metastasis.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2001.11a
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• pp.106-106
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• 2001

Matrix metalloproteases (MMPs) 는 다양한 세포에서 전이와 침윤성에 중요한 역할을 한다. MMP의 내인성 저해제인 tissue inhibitor of motalloprotease-2 (TIMP-2) 는 MMP-2에 높은 특이성을 지닌다. MMP-2와 TIMP-2사이의 불균형은 침윤성과 전이와 같은 병리학적 과정과 관계되는 extracellular matrix (ECM)의 퇴화를 일으킨다. TIMPs는 분비되는 분자이기 때문에 특정한 암의 유전자 치료에 사용될 가능성을 지닌다. 본 연구에서는 MMP-2가 H-ras에 의해 유도된 침윤성에 책임지는 것으로 보여지는 H-ras MCF10A 세포에 TIMP-2 유전자를 함유하는 retrovirus를 이용하여 연구하였다. TIMP-2 유전자를 함유하는 재조합 retrovirus는 PG13 세포를 infection 시키는데 사용되었다. H-ras MCF10A 세포는 PGl3 세포의 conditioned media로 처리되었을 때, gelatin zymography에서 MMP-2의 분비가 농도의존적으로 저해되었다. 또한 retrovirus에 의한 TIMP-2의 과잉 발현은 농도의존적으로 H-ras MCF10A 세포의 침윤성과 이동성을 상당히 감소시킨다. 이와 같은 실험 결과는 TIMP-2가 H-ras MCF10A 세포에서 MMP-2 분비와 세포의 침윤성, 이동성을 감소시키는 역할을 지닌다는 것과 TIMP-2 유전자를 함유하는 retrovirus가 효과적으로 MMP-2 분비, 세포 침윤성, 세포 이동성을 감소시켰다는 것을 보여 준다. 이는 암의 예방과 치료를 위한 유전자 치료법의 적용에 상당한 가능성을 제시한다.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2016.05a
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• pp.711-714
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• 2016

Recombinant baculovirus was reconstructed with useful genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). This reconstructed vector was infected into various cell lines and tissues. We investigated gene transfer and gene expression of this reconstructed vector in comparison to other vectors and recognized that this reconstructed vector was higher effective than any other control vector.

• Korean Journal of Weed Science
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• v.32 no.1
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• pp.10-16
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• 2012

This study was carried out to investigate the agronomic traits, comparison of weed characteristics and possibility of gene flow in 'vitamin A enforced GM rice' and the donor plant, 'Nagdong'. The GM rice was not significantly different agronomic traits compared to the donor plant, Nagdong. Weed population changes were investigated in the cultivation of the GM rice and the donor plant, Nagdong. Dominant weed species and their dry matter did not show the difference between GM rice and the donor plant, Nagdong in macro-GM crop field. Dominant weed species with the GM rice and the donor plant, Nagdong were Monochoria vaginalis, followed by Eleocharis kuroguwai, Echinochloa crus-galli and Lindernia procumbens. The detection of gene from the GM rice was done using PCR, gene flow can't be detected by weed species. Results of this study on the agronomic traits, weed characteristics and possibility of gene flow has elucidated that GM rice might not be different from the donor plant, Nagdong.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.171-180
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• 2001

Compared to other gene transfer system, the advantages of retrovirus-mediated gene transfer are technical ease, efficient expression and genetic stability. Despite the high potency of the retrovirus vector system in gene transfer, one of the drawbacks is a difficulty in concentration of virus stock. To overcome this problem, we tested a new retrovirus vector system producing the progeny retrovirus particles encapsidated with VSV-G (vesicular stomatitis virus G glycoprotein). The infectivity of this virus was not sacrificed by ultracentrifugal concentration and the host cell range extended from all mammalian to fish embryos. Virus titer after 1,000 x concentration was more than 10$^{8}$ CFU/ $m\ell$ on most of the target cell lines. We applied this pantropic viruses in transgenic chicken production by injecting the concentrated (100$\times$) stock into subgerminal cavity of stage X chicken embryos. The survival rate of chicken embryos after injection was about 20% and gene integration rate in surviving embryos was scored almost 100%. Analyses of RT-PCR and fluorescence microscopy, however, showed no evidence of the transgene expression.

• Korean Journal of Microbiology
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• v.31 no.4
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• pp.286-291
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• 1993

The $Km^{r}$ gene and plasmid of natural isolate and genetically modified microorganisms (GMM) rearranged by conjugation in water environments were comparatively analyzed by agarose gel electrophoresis and Southern analysis. The transfer rates of the $Km^{r}$ gene from GMM strains were generally 100 times higher than thosc of natural iso]ate(DKI) under laboratory cnvironments, but their transfer rate was not much different in Moosimcheon River water. The conjugants obtained in LB(Luria-Bertani broth) and FW(filtered river water) water under laboratory conditions showed same number of the plasmids. but the sizes of the plasmids were changed. The $Km^{r}$ gene in the conjugants was found in the same position as the pDKJO] $Km^{r}$ plasmid. In case of the GMM strains as donor. the large plasmids of 180 kb appeared in conjugants obtained in LB and FW water. Especially, the $Km^{r}$ gene in the donor of DKC600 was found to be inserted into chromosome of the conjugant obtained in FW water. However. in the conjugants obtained from DKl and DKB 701 in Moosimcheon River water, the plasmids were rearranged by 4 and 8. respectively, and all of them showed hybridization by the $Km^{r}$ probe. But the small plasmids of the recipient disappeared in the conjugant from DKC600 as donor, and the rearranged plasm ids and chromosome in the conjugants were observed to be hybridized with the $Km^{r}$ probe. Therefore, rearrangement of $Km^{r}$ gene and plasmids by conjugation was found to be afTected diversely by cellular characteristics as well as by environmental factors.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.9-14
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• 2003

The pBT-L transgenic mice carrying human TPO gene in conjunction with bovine $\beta$-casein promoter express human TPO in milk during lactation. In this study, stability of germ line transmission and expression of pBT-L transgene integrated into host chromosome were monitored up to generation F10 of transgenic pBT-L/15 line. When male mouse of generation F8 was crossbred with normal females, approximately half of offsprings (51.3$\pm$18.98%) were identified as transgenic mice. Generation F9 and F 10 mice also showed similar transmission rates (43.8$\pm$18.98% and 71.4$\pm$26.98%, respectively), implying that pBT-L transgene can be transmitted stably up to long term generation in the transgenic mice. Expression levels of human TPO from milk of generation F9 and F10 mice were 1.1$\pm$0.33 mg/ml and 1.1 $\pm$0.45 mg/ml, respectively, which are similar to expression level of generation F2 mice. In conclusion, our results suggest that transgenic animals once established will continuously pass their transgenes to the progeny through the breeding program with the same productivity of human protein in their milk.

• Journal of Life Science
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• v.21 no.4
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• pp.529-533
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• 2011

Metallothioneins (MT) are a group of low-molecular weight, cysteine-rich, intracellular proteins that are encoded by a family of genes containing at least 10 functional isoforms in human. The expression and induction of these proteins is associated with protection against DNA damage, oxidative stress, and apoptosis. Many studies have shown increased expression of MT in various human tumors, whereas MT is down-regulated in certain tumors such as hepatocellular carcinoma and liver adenocarcinoma. Hence, the expression of MT is not universal to all human tumors but may depend on the differentiation status and proliferative index of tumors, along with other tissue factors and gene mutations. Using Northern blot analysis, we found that laminin induced expression of MT-1 in HSG and PC12 cells, which can be differentiated by laminin, but had no effect on MB-231, MDA-435, and PC-3 cells, which cannot be differentiated by laminin. In addition, we analyzed the expression level of the MT-1 gene in five prostate cancer cell lines possessing different metastatic potential. The expression of MT-1 in normal and less malignant cells (RWPE-1 and WPE1-NA22) was high and up-regulated by laminin, whereas the expression of MT-1 in WPE1-NB14, WPE1-NB11, and WPE1-NB26 cells (malignant) was extremely low and not elevated by laminin. These results suggest that the MT-1 gene is involved in laminin-mediated differentiation and affects the metastatic potential of tumor cells.

• Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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• 2015.10a
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• pp.838-841
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• 2015

Recombinant baculovirus vector is composed of genes of polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), and protein transduction domain (PTD). This recombinant baculovirus vector was transfected into cell lines and tissues and then found out a novel possibility in view of gene transfer and gene expression in comparison to other vector systems. Efficacy of gene transfer and gene expression of this recombinant baculovirus vector was higher than any other vector system.