• 제목/요약/키워드: gene delivery

검색결과 329건 처리시간 0.038초

Screening of silkworm strains for efficient recombinant protein production by Autographa californica nucleopolyhedrosis virus (AcNPV)

  • Park, Yoon Mi;Kim, Kyung A;Kang, Min Uk;Park, Kwan Ho;Nho, Si Kab
    • International Journal of Industrial Entomology and Biomaterials
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    • 제28권1호
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    • pp.10-18
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    • 2014
  • Baculoviruses base vectors come to be regarded as methods for in vivo gene delivery and transient expression to the silkworm. In the case of silkworm, B. mori, two types of baculoviruses, AcNPV (Autographa californica nuclear polyhedrosis virus) and BmNPV (Bombyx mori nuclear polyhedrosis virus), are potentially applicable as vectors. Recently, AcNPV showed promising results with some silkworm strains despite different host-specificities. We searched for a highly-permissive silkworm strain in the B. mori stocks of Kyungpook National University that could produce high levels of recombinant protein. Seventy strains were screened using the recombinant AcNPV/BmA3-Luc virus. Based on the measured luciferase activity, the strains could be divided into three groups, high-, middle-, and low-permissive strains, according to their relative recombinant protein expression levels. At 48 hours post-injection, the luciferase activity in the high-permissive strains was 500-fold greater than that of the low-permissive strains. At 72 hours post-injection, a significant elevation in luciferase activity was observed in the hemocytes of all strains. Then, based on the above results, the High Permissive Strain (HPS) S10 and the Low Permissive Strain (LPS) S39 were pick up and was carried out Dot blotting, RT-PCR and Real time PCR.

Inhibition of Inducible Nitric Oxide Synthase and Cyclooxygenase-2 by Gamijihwang-tang Via Suppression of Nuclear Factor-B Activation in RAW 264.7 cells

  • Jang Du-Hyun;Kim Ji-Young;Han Eun-Hee;Park Hee-Ok;Kim Dong-Hee;Jeong Hye-Gwang;Yoo Dong-Yeol
    • 동의생리병리학회지
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    • 제19권5호
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    • pp.1405-1410
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    • 2005
  • Asthma is recognized today as an inflammatory disease of the lung characterized by acute non-specific airway hypersensitiveness in association with chronic pulmonary inflammation. Gamijihwang-tang(GJT), a fortified prescription of YMJHT, is applied for the treatments of chronic coughing and asthma, and post-delivery coughing and asthma in the gynecology. Also in the clinical practice, GJT is known to be very effective for controlling coughing and asthma as a cold sequoia. In this study, we investigated the effects of GJT on the lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) production, and on the level of inducible nitric oxide synthase (iNOS) and Cyclooxygenase-2 expression in murine macrophage RAW 264.7 cells. We found that GJT inhibited LPS-induced NO and $PGE_2$ production in a dose dependent manner. Furthermore, GJT inhibited the expression of LPS-induced iNOS and COX-2 protein and mRNA expression in RAW 264.7 macrophages. Treatment with GJT of RAW 264.7 cells transfected with a reporter construct indicated a reduced level of LPS-induced nuclear factor-KB (NF-kB) activity and effectively lowered NF-kB binding as measured by transient transfection assay. These results suggest that the main inhibitory mechanism of the GJT may be the reduction of iNOS and COX-2 gene expression through blocking of NF-kB activation.

Somatic embryogenesis from the axillary meristems of peanut (Arachis hypogaea L.)

  • Singh, Shweta;Hazra, Sulekha
    • Plant Biotechnology Reports
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    • 제3권4호
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    • pp.333-340
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    • 2009
  • Developmental anomalies in the plumule meristem of peanut (Arachis hypogaea L.) somatic embryos resulted in poor shoot differentiation and reduced plant recovery. Existing meristems with caulogenic potential have never been tested for embryogenesis in peanut. The present experiment was designed to test the mature zygotic embryo axis derived plumule with three meristems for somatic embryogenesis. Embryogenic masses and embryos developed from the caulogenic meristems in the axils. Exposure of 2 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation temporarily which then regained the ability to form the shoot on withdrawal of 2,4-D. Exposure of 4 weeks in primary medium with $90.5{\mu}M$ 2,4-D suppressed the shoot tip differentiation irreversibly. No shoot formation was noted from the tips in any of the cultures which were in secondary medium with $13.6{\mu}M$ 2,4-D. Development of somatic embryos directly from axillary meristems was confirmed histologically. Conversion frequency of these embryos was 11%. Thus, in this report, we describe a method to obtain somatic embryos from the determined organogenic buds of the axillary meristem, by culturing the nodal explant vertically on embryo induction medium. It also displays the possibility of obtaining both embryogenic and organogenic potential in two parts of the same explant simultaneously. The possibility of extending this approach for genetic transformation in in vivo system through direct DNA delivery or Agrobacterium injection in meristems can also be explored. Using Agrobacterium rhizogenes, we have demonstrated the possibility of gene transfer in the axillary meristems of seed-derived plumule explant.

싸움소의 증례를 통한 동물복지형 헤드기어의 개발 (Development of animal welfare type headgear in the case of fighting bulls)

  • 김택석;김충희
    • 한국동물위생학회지
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    • 제38권4호
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    • pp.253-257
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    • 2015
  • Bull-boxing in South Korea is a form of competitions in which bulls measure their strength with each other in the form of pushing by putting their heads together. Bull-boxing has been admitted after being designated as a play culture by the government because it does not cause great injuries to animals' bodies and has strong positive aspects such as contributing to livestock farmers' leisure activities, encouragement of animal husbandry, and communities' economic development. However, bull-boxing sometimes causes damage to the head and horns due to the heavy bodies. According to the results of examinations of these damage cases, damage that caused irregular shapes of the base of horns was identified in 31.5% of fighting bulls. The damage to horns is thought to have been caused by the great forces of fighting bulls repeatedly imposed to each other's horns during bull-boxing that caused minute fractures leading to damage to the blood vessels inside the horns resulting in the blocking of delivery of nutrients to the horns causing the deformation of the horns into abnormal shapes. Since bulls' bodies are injured during bull-boxing although the injuries are small as bull-boxing is mainly conducted by pushing, animal protection groups regard bull-boxing as cruelty to animals and request to stop bull-boxing. The present study aims to develop animal welfare type headgears that can protect bulls' horns and heads in order to protect fighting bulls during bull-boxing in terms of animal protection and welfare.

Effect of Drug Loading on the Physicochemical Properties and Stability of Cationic Lipid-based Plasmid DNA Complexes

  • Jeong, Ui-Hyeon;Jung, Ji-Hye;Davaa, Enkhzaya;Park, Se-Jin;Myung, Chang-Seon;Park, Jeong-Sook
    • Journal of Pharmaceutical Investigation
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    • 제39권5호
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    • pp.339-343
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    • 2009
  • Recently, co-delivery of drug and gene has been attempted for higher therapeutic effects of anticancer agents. In this study, cationic liposomes were prepared using 1,2-dioleoyl-3-trimethylammoniopropane (DOTAP) as a cationic lipid to investigate the effect of drug loading on the physicochemical characteristics of cationic liposomes/DNA complexes. The complex formation between cationic liposomes and negatively charged plasmid DNA was confirmed and the protection from DNase was observed. Particle size of complexes was reduced not by drug loading, but by the increased ratio of cationic lipid to plasmid DNA. Meanwhile, zeta potential of complex was increased by the addition of cationic liposomes to complexes and the effect of drug loading on the zeta potential was not much higher than on particle size. Gel retardation of complexes was indicated when the complexation weight ratios of cationic lipid to plasmid DNA were higher than 24:1 for drug free complexes and 20:1 for drug loaded ones, respectively. Agarose gel retardation showed the similar complexation between plasmid DNA and drug free liposomes or drug loaded liposomes. Both complexes protected plasmid DNA from DNase independent of complexation temperature. From the results, drug loading may affect not the complex formation of cationic liposomes and plasmid DNA, but the particle size of complex.

Altered Gene Expression of Caspase-10, Death Receptor-3 and IGFBP-3 in Preeclamptic Placentas

  • Han, Jae Yoon;Kim, Yoon Sook;Cho, Gyeong Jae;Roh, Gu Seob;Kim, Hyun Joon;Choi, Won Jun;Paik, Won Young;Rho, Gyu Jin;Kang, Sang Soo;Choi, Wan Sung
    • Molecules and Cells
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    • 제22권2호
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    • pp.168-174
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    • 2006
  • Enhanced apoptosis has been observed in the placentas of women with preeclampsia, but few studies have examined changes at the molecular level. This study was designed to detect genes specifically expressed in full-term preeclamptic placentas. Tissue samples were collected immediately after cesarean delivery from 11 normal and 8 preeclamptic placentas at 35-40 weeks of gestation. Total RNAs were extracted and hybridized to a cDNA microarray. Results were confirmed by reverse-transcription polymerase chain reaction (RT-PCR), Western blotting and immunohistochemistry. Hematoxylin and eosin and TUNEL staining were also performed to confirm apoptosis in preeclamptic placentas. Among 205 genes, three were up- or downregulated in preeclamptic placentas. The expression of caspase-10 and death receptor 3 (DR-3) was significantly increased, whereas insulin-like growth factor binding protein-3 (IGFBP-3) was strongly downregulated. RT-PCR analysis and Western blotting confirmed these effects. Immunohistochemical analysis showed that the DR-3, caspase-10 and IGFBP-3 proteins were localized in the syncytial membrane. Apoptosis in the trophoblast was also increased in term placentas from women with pregnancies complicated by preeclampsia. These results suggest that caspase-10, DR-3 and IGFBP-3 are involved in apoptosis in the preeclamptic placenta.

비선형회귀 확산모형을 이용한 반도체 시장수요 추정 (Estimation of Semiconductor Market, Using NLS Diffusion Model)

  • 김진;고경일
    • 디지털융복합연구
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    • 제12권3호
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    • pp.141-147
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    • 2014
  • 확산모형은 시장의 예측이나 그 방법론의 연구를 위해 마케팅에서 광범위하게 다뤄졌을 뿐 아니라, 경제학에서도 다양하게 활용되어 왔다. 특히 Bass 모형은 Rogers의 혁신확산 및 수명주기이론을 간단한 수리적 모형으로 표현할 수 있기에 혁신적 신제품의 채택과 확산을 설명하는데 널리 활용되었다. 그럼에도 불구, 확산모형은 '산업의 쌀' 이라 일컬어지는 반도체의 수요예측에 일부 선도적 연구를 제외하고는 활용된 바 없다. 이에 Bass 모형에서 진일보한 비선형회귀 접근법 확산모형을 활용, 전력반도체 중 전기전자기구의 필수 스위치로 채택되는 MOSFET의 수요를 추정하여 수명주기를 예측하고 그 과정을 설명함으로써 산업관계자는 물론 반도체와 전기전자산업 정책입안자에게 중요한 시사점을 전달하고자 한다.

Overexpression of microRNA-612 Restrains the Growth, Invasion, and Tumorigenesis of Melanoma Cells by Targeting Espin

  • Zhu, Ying;Zhang, Hao-liang;Wang, Qi-ying;Chen, Min-jing;Liu, Lin-bo
    • Molecules and Cells
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    • 제41권2호
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    • pp.119-126
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    • 2018
  • microRNA (miR)-612 shows anticancer activity in several types of cancers, yet its function in melanoma is still unclear. This study was undertaken to investigate the expression of miR-612 and its biological relevance in melanoma cell growth, invasion, and tumorigenesis. The expression and prognostic significance of miR-612 in melanoma were examined. The effects of miR-612 overexpression on cell proliferation, colony formation, tumorigenesis, and invasion were determined. Rescue experiments were conducted to identify the functional target gene(s) of miR-612. miR-612 was significantly downregulated in melanoma tissues compared to adjacent normal tissues. Low miR-612 expression was significantly associated with melanoma thickness, lymph node metastasis, and shorter overall, and disease-free survival of patients. Overexpression of miR-612 significantly decreased cell proliferation, colony formation, and invasion of SK-MEL-28 and A375 melanoma cells. In vivo tumorigenic studies confirmed that miR-612 overexpression retarded the growth of A375 xenograft tumors, which was coupled with a decline in the percentage of Ki-67-positive proliferating cells. Mechanistically, miR-612 targeted Espin in melanoma cells. Overexpression of Espin counteracted the suppressive effects of miR-612 on melanoma cell proliferation, invasion, and tumorigenesis. A significant inverse correlation (r = -0.376, P = 0.018) was observed between miR-612 and Espin protein expression in melanoma tissues. In addition, overexpression of miR-612 and knockdown of Espin significantly increased the sensitivity of melanoma cells to doxorubicin. Collectively, miR-612 suppresses the aggressive phenotype of melanoma cells through downregulation of Espin. Delivery of miR-612 may represent a novel therapeutic strategy against melanoma.

DNA가 봉입된 Poly(D,L-lactic-co-glycolic acid) 미립구의 제조 및 시험관내 방출 (Preparation and In Vitro Release of DNA-Loaded Poly(D,L-lactic-co-glycolic acid) Microspheres)

  • 손혜정;김진석
    • 폴리머
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    • 제29권1호
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    • pp.69-73
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    • 2005
  • 비바이러스성 유전자 전달체의 주요 단점인 낮은 transfection 효율에 기인한 반복투여 등을 극복하기 위하여 poly (D,L-lactide-co-glycolide)를 이용하여 DNA가 봉입된 미립구를 제조하였다. pDNA 그 자체 또는 여러 비율의 키토산/pDNA 복합체를 사용하여 봉입하였고, 그 결과 44%(pDNA 그 자체), 5%(0.7:1 미토산/pDNA 복합체), 그리고 8%(1:1 키토산/pDNA 복합체)의 봉입효율을 나타내었다. 주사전자현미경(SEM)을 통해 본 표면구조에서는 미립구 제조 직후에서는 매우 매끈한 구형을 보이다가 제조 후 41일 경에는 찌그러진 다공성의 구조를 보였는데 이는 미립구 제조에 사용한 poly(D,L-lactic-co-glycolic acid)(PLGA) 고분자의 분해에 의한 것으로 생각된다. 시험관내 방출실험에서는 0.7:1 키토산/pDNA 복합체를 사용한 미립구에서 47%의 pDNA가 26일만에 방출된데 반해, pDNA 그 자체 혹은 1:1 키토산/pDNA 복합체를 사용한 미립구에서는 각각 15% 혹은 32%의 pDNA 방출을 나타내었다.

Nitric oxide-Releasing Chitosan Nanoparticles; A Potential Impeding Strategy Against Salinity Stress in Arabidopsis thaliana

  • Waqas Rahim;Anjali Pande;Nusrat Jahan Methela;Da-Sol Lee;Bong-Gyu Mun;Hak-Yoon Kim;Byung-Wook Yun
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2022년도 추계학술대회
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    • pp.157-157
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    • 2022
  • Plants being sessile are prone to various abiotic challenges, including salinity. Plants generally cope with salt stress by regulating their endogenous NO levels. NO exogenously applied in various forms also successfully impedes the salt stress, but its small size, short half life, and high volatility rate hamper its application in agriculture. NO application via CS as a nanocarrier is an alternate option to ensure the optimal kinetic release of NO for a long period compared to the free NO form. Herein, we synthesized and characterized GSNO-CS NP by ionic gelation of TPP with CS and then reacting with GSH, followed by reaction with NaNO2 suspension. The synthesized NPs were characterized using non-destructive analytical techniques such as DLS, FTIR, and SEM to ensure their synthesis and surface morphology. NO-release profile confirmed optimal kinetic NO release for 24 h from NO-CS NP as compared to free NO form. The efficiency of NO-CS NP was checked on Arabidopsis plants under salinity stress by gauging the morphological, physiological, and enzymatic antioxidant system and SOS pathway gene expression levels. Overall, the results revealed that NO-CS NP successfully mitigates salinity stress compared to free GSNO. Concluding, the findings provide sufficient experimental evidence for the application of nanotechnology to enhance NO delivery, thus inducing more benefits for the plants under stress conditions by mitigating the deleterious impacts of salt stress on the morphological and physiological status of the plants, and regulating the ions exchange by overexpression of SOS pathway candidate genes.

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