• Title/Summary/Keyword: transfection efficiency

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Cationic Emulsions with Galactosylated Chitosan as a Novel Gene Delivery System

  • Choi, Woo-Jeong;Choi, Sung-Hee;Ahn, Woong-Shick;Kim, Chong-Kook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.427.1-427.1
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    • 2002
  • To improve stability and transfection efficiency, a novel combination of cationic emulsion and galactosylated chitosan was developed for targeted gene delivery. Six formulations of cationic liposome and our novel emulsion were prepared for comparison of stability and transfection efficiency. Cationic liposomes composed of 3[N-(N.N dimethylaminoethylene) carbamoyl] cholesterol (DC-Chol) and dioleyl phophatidyl ethanolamine (DOPE) were prepared by extrusion method and cationic emulsions composed of DC-Chol. DOPE. castor oil, and Tween 80 were prepared by sonication method. (omitted)

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Effect of Polyethylenimine Type in Polylactic acid Nanoparticles/DNA Complex on the Transfection Efficiency

  • Chae, Jong-Hyuck;Park, Yu-Mi;Kim, Kyeong-Man;Lee, Yong-Bok;Shin, Sang-Chul;Oh, In-Joon
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.425.1-425.1
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    • 2002
  • Nanoparticles of polylactic acid (PLA) and polyethylenimine (PEI) as an effective gene delivery agent were prepared and characterized. As a model plamid DNA. PME185/$\beta$-gal. a mammalian expression vector. and fluorescence enhancing protein (pEGHP) were used. The effects of PEI type on the physical properties of nanoparticles and transfection efficiency were examined. (omitted)

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Nonionic Amphiphilic Surfactant Conjuncted Polyethyleneimine as a New and Highly Efficient Non-viral Gene Carrier

  • Yin, Dongfeng;Chu, Cang;Ding, Xueying;Gao, Jing;Zou, Hao;Gao, Shen
    • Macromolecular Research
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    • v.17 no.1
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    • pp.19-25
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    • 2009
  • In order to enhance the gene delivery efficiency and decrease the cytotoxicity of polyplexes, we synthesized Solutol-g-PEI by conjugating polyethyleneimine (PEI) to Solutol (polyoxyethylene (10) stearate), and evaluated its efficiency as a possible nonviral gene carrier candidate. Structural analysis of synthesized polymer was performed by using $^1H$-NMR. Gel retardation assay, particle sizes and zeta potential measurement confirmed that the new gene carrier formed a compact complex with plasmid DNA. The complexes were smaller than 150 nm, which implicated its potential for intracellular delivery. It showed lower cytotoxicity in three different cell lines (Hela, MCF-7, and HepG2) than PEI 25 kDa. pGL3-lus was used as a reporter gene, and the transfection efficiency was in vitro measured in Hela cells. Solutol-g-PEI showed much higher transfection efficiency than unmodified PEI 25 kDa.

Examination of alginate/PEl/DNA polyplex as a gene delivery system: enhancing transfection efficiency in the presence of serum and reducing cytotoxicity

  • Jiang, Ge;Min, Sang;Kim, Mi-Na;Lim, Mi-Jung;Yeom, Young-Il
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.277.2-278
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    • 2002
  • Synthetic vectors have been considered as a safer and more versatile alternative to viral-based gene delivery systems. A variety of simple synthetic vector systems such as cationic lipid- and polymer-complexed plasmid DNA were shown to have a significant transfection activity in vitro but their use in vivo has been hampered by the decrease in transfection efficiency mediated by non-specific electrostatic interactions with serum components. (omitted)

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Enhancement of Emulsion-mediated Gene Expression by Using Chitosan as a Pre-Condensing Agent

  • Chun, Soo-Kyung;Choi, Sung-Hee;Kim, Adele;Ahn, Woong-Shick;Kim, Chong-Kook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.426.3-427
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    • 2002
  • The aim of this study was to enhance the transfection efficiency of emulsion-mediated gene expression by using chitosan, Conventional DNA/emulsion complexes and precondensed DNA/emulsion complexes were prepared by adding either naked or precondensed plasmids to cationic emulsion. The zeta potential. TEM, and size of transfection complexes were measured. In vitro transfection efficiency for boty complexes was also studied by several methods: flow cytometer, expression analysis by confocal microscope, RT-PCR, and in addition. (omitted)

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R3V6 Amphiphilic Peptide with High Mobility Group Box 1A Domain as an Efficient Carrier for Gene Delivery

  • Ryu, Jaehwan;Jeon, Pureum;Lee, Minhyung
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3665-3670
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    • 2013
  • The R3V6 peptide includes a hydrophilic arginine stretch and a hydrophobic valine stretch. In previous studies, the R3V6 peptide was evaluated as a gene carrier and was found to have low cytotoxicity. However, the transfection efficiency of R3V6 was lower than that of poly-L-lysine (PLL) in N2A neuroblastoma cells. In this study, the transfection efficiency of R3V6 was improved in combination with high mobility group box 1A domain (HMGA). HMGA is originated from the nuclear protein and has many positively-charged amino acids. Therefore, HMGA binds to DNA via charge interaction. In addition, HMGA has a nuclear localization signal peptide and may increase the delivery efficiency of DNA into the nucleus. The ternary complex with HMGA, R3V6, and DNA was prepared and evaluated as a gene carrier. First, the HMGA/DNA complex was prepared with a negative surface charge. Then, R3V6 was added to the complex to coat the negative charges of the HMGA/DNA complex, forming the ternary complex of HMGA, R3V6, and DNA. A physical characterization study showed that the ternary complex was more stable than the PLL/DNA complex. The HMGA/R3V6/DNA complex had a higher transfection efficiency than the PLL/DNA, HMGA/DNA, or R3V6/DNA complexes in N2A cells. Furthermore, the HMGA/R3V6/DNA complex was not toxic to cells. Therefore, the HMGA/R3V6/DNA complex may be a useful gene delivery carrier.

STABLE TRANSFORMATION OF CULTURED CHICKEN CELLS

  • Han, J.Y.;Shin, Y.S.;Shoffner, R.N.;Guise, K.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.4
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    • pp.581-589
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    • 1993
  • A plasmid vector, $RSVLTR/{\beta}G2$, containing lacZ gene under the control of the RSVLTR promoter were transfected into chicken embryo fibroblasts by three different transfection methods. Calcium phosphate, lipsome and DEAE-dextran techniques were applied for transfection of chicken cells. A histochemical assay with X-gal was used as a simple method for screening transfected cells. Plasmid $RSVLTR/{\beta}G2$ was expressed proficiently in the chicken embryo fibroblast. Calcium phosphate-DNA precipitate transfection resulted in the highest efficiency for transient expression of $RSVLTR/{\beta}G2$. Transfected cells formed colonies on the 9th day of incubation indicating stable transformation of the inserted plasmid.

Single-cell Electroporation and Gene Transfection using MEMS-based Microdevice with Cantilever-type Microelectrode (멤스 기반의 캔틸레버 형 전극을 가진 마이크로 디바이스를 이용한 단일세포의 Electroporation 및 유전자 Transfection)

  • Cho, Young-Hak;Kim, Beom-Joon
    • Journal of the Korean Society for Precision Engineering
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    • v.27 no.5
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    • pp.85-91
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    • 2010
  • In this paper, we present details on fabrication of single-cell electroporation microdevice, practical experiments of single-cell electroporation with our fabricated microdevice. Also, the continuous electroporation for the continuous flow of cells is used for high-throughput electroporation. The delivery efficiency and cell viability tests are provided and the successful GFP transfection into cells is also evaluated with a fluorescent microscope after electroporation. This device enables to reduce the size of samples and thus the use of small amount of reagents. Also, it makes it possible to permit to avoid cell discrimination (transfected cells versus non-transfected cells) encountered when traditional bulk electroporation is held.