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Results 651 - 700 of 1865
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Hansen Wiebke - - 2005
Retroviral vectors released from mouse-derived packaging cell lines are inactivated in human sera by naturally occurring antibodies due to the recognition of Galalpha1,3Galbeta1,4GlcNAc (alphagal-epitope) decorated surface proteins. In this study, an extensive analysis of the glycosylation potential of NIH3T3-derived PA317 packaging cells using combined MALDI/TOF-MS and HPAE-PAD reveals that 34% ...
Qin Jian-Min - - 2005
BACKGROUND: Signal regulatory protein alpha1 (Sirpalpha1) is a negative regulatory factor, and inhibits receptor tyrosine kinase-dependent cell proliferating signal. This study was undertaken to observe the effect of signal regulatory proteinalpha1 (Sirpalpha1)on gankyrin,cyclin D1,CDK4 and Fas expression in Sk-hep1 mouse hepatoma carcinoma cell line. METHODS: BOSC 23 packed cells were ...
Zhai Ling - - 2005
In contrast to extensive studies on the roles of molecular chaperones, such as heat shock proteins, there are only a few reports about the roles of GRP78/BiP, an endoplasmic reticulum (ER) stress-induced molecular chaperone, in mammalian cell responses to DNA-damaging stresses. To investigate whether GRP78/BiP is involved in resistance to ...
Alokail Majed S - - 2005
Epidermal growth factor receptor (EGFR) is activated by autocrine growth factors in many types of tumours, including breast tumours. This receptor has been linked to a poor prognosis in breast cancer and may promote proliferation, migration, invasion, and cell survival as well as inhibition of apoptosis. Human breast ductal carcinoma ...
Singh Balraj - - 2005
Cyclooxygenase-2 (COX-2), an inducible enzyme involved in prostaglandin (including PGE(2)) biosynthesis, is overexpressed in several epithelial malignancies including breast cancer. We tested the hypothesis that COX-2 overexpression in breast cancer cells results in increased cell motility and invasion. COX-2 overproducing cells were generated by stable transfection of several human breast ...
Thomas Philip P Department of Pharmacology, University College London, Gower Street, London, WC1E 6BT, - - 2005
The HEK cell line has been extensively used as an expression tool for recombinant proteins since it was generated over 25 years ago. Although of epithelial origin, its biochemical machinery is capable of carrying out most of the post-translational folding and processing required to generate functional, mature protein from a ...
Morisaki Kuniaki - - 2005
Single nucleotide polymorphism (SNP) analyses of the ABCG2 gene have revealed three nonsynonymous SNPs resulting in the amino acid changes at V12M, Q141K and D620N. To determine whether the SNPs have an effect on drug transport, human embryonic kidney cells (HEK-293) were stably transfected with full length ABCG2 coding wild-type ...
Gao Junwei - - 2005
Aquaporin (AQP) is a kind of channel-forming membrane glycoprotein that mediates osmotic water transport. The present study aimed to establish a cell line stably transfected with AQP1 to measure osmotic water permeability. The recombinant plasmid was constructed by subcloning the full-length rat AQP1 cDNA into pEGFP-C3 vector, named pEGFP/AQP1. Human ...
Feng De-Yun - - 2005
AIM: To study the effect of hepatitis C virus nonstructural region 3 (HCV NS3) protein on proliferation and transformation of normal human liver cell line. METHODS: QSG7701 cells were transfected with pRcHCNS3-5', pRcHCNS3-3' and pRcCMV using lipofectamine transfecting technique and selected with G418 method. Expression of HCV NS3 protein was ...
Prasad Tekkatte Krishnamurthy - - 2005
Variation in transfection efficiency observed in different cell-types is poorly understood. To investigate the influence of endocytic activity on lipid-mediated transfections, we have monitored both the processes in 12 different cell-types. The endocytic activity shows a strong positive correlation (P < 0.01), with transfection efficiency. Treatment with wortmannin resulted in ...
Poliseno Laura - - 2005
Testing of short interfering RNAs (siRNAs) to knock-down gene expression is complicated in primary differentiated cells due to their limited availability and short in vitro life span. The objective of this study was to bypass these limitations by testing selected siRNAs in a heterologous cell line. A plasmid containing a ...
Warmann Steven W - - 2005
BACKGROUND/PURPOSE: Enhanced green fluorescent protein (eGFP) is widely used as a marker in different biologic systems. After vector transfection, eGFP is expressed by eukaryotic cells and can be visualized using fluorescent microscopy. The aim of this study was to establish an eGFP-transfected human hepatoblastoma (HB) cell line as tool for ...
Nicholson Linda J - - 2005
The success of RNA interference (RNAi) in mammalian cells, mediated by siRNAs or shRNA-generating plasmids, is dependent, to an extent, upon transfection efficiency. This is a particular problem with primary cells, which are often difficult to transfect using cationic lipid vehicles. Effective RNAi in primary cells is thus best achieved ...
Männistö Marjo - - 2005
BACKGROUND: Retinal pigment epithelium (RPE) maintains the function of photoreceptors and eyesight and is an important target for gene delivery. Since in some diseases RPE cells proliferate uncontrollably, we investigated the role of cell cycle in non-viral gene delivery into a RPE cell line (D407). METHODS: D407 (human) cells were ...
Funnell Barbara E - - 2005
Plasmid partition systems are essential for the stability and thus the survival of low-copy-number plasmids in growing bacterial populations. The partition reaction is responsible for proper intracellular distribution of plasmids in the bacterial cell cycle. One common step in most partition models is the pairing of plasmids to each other ...
Malfatti Michael A - - 2005
UDP-glucuronosyltransferase proteins (UGT) catalyze the glucuronidation of both endogenous and xenobiotic compounds. In previous studies, UGT1A1 has been implicated in the detoxification of certain food-borne carcinogenic-heterocyclic amines. To determine the importance of UDP-glucuronosyltransferase 1A1 (UGT1A1) in the biotransformation of the cooked-food carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), genetically modified CHO cells that are ...
Cheng Sheng-Quan - - 2005
AIM: To investigate the survivin gene expression in human hepatocellular carcinoma cell line SMMC-7721 and the effects of survivin gene RNA interference (RNAi) on cell apoptosis and biological behaviors of SMMC-7721 cells. METHODS: Eukaryotic expression vector of survivin gene RNAi and recombinant plasmid pSuppressorNeo-survivin (pSuNeo-SVV), were constructed by ligating into ...
Watanabe Hisashi - - 2005
Hypophosphatasia (HOPS) is a clinically heterogeneous heritable disorder characterized by defective skeletal mineralization, deficiency of tissue-nonspecific alkaline phosphatase (TNSALP) activity, and premature loss of deciduous teeth. To date, various mutations in the TNSALP gene have been identified. Especially, A115V located in exon 5 has been detected in a Japanese patient ...
Kamimura Yutaka - - 2005
alpha2,8-Sialyltransferase (alpha2,8S-T, GD3 synthase) has been reported to be involved in the enhanced cell proliferation of malignant tumors. Using a cloned cDNA of alpha2,8S-T, transfectant cells were established and the effects of the gene expression on the cell phenotypes were analyzed. In contrast with PC12 cells, in which we reported ...
Paterson L - - 2005
The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power ...
Xiao De-Sheng - - 2005
AIM: To investigate the effect of deleted pancreatic cancer locus 4 (DPC4) gene transfection on biological behaviors of human colorectal carcinoma cells and the role of DPC4 gene in colorectal carcinogenesis. METHODS: PcDNA3.1-DPC4 plasmid was re-constructed by gene-recombination technology. SW620 cells, a human colorectal carcinoma cell line, were transfected with ...
Isalan Mark - - 2005
High-throughput gene analysis would benefit from new approaches for delivering DNA or RNA into cells. Here we describe a simple system that allows any molecular biology laboratory to carry out multiple, parallel cell transfections on microscope coverslip arrays. By using magnetically defined positions and PCR product-coated paramagnetic beads, we achieved ...
Hu Zhong-Liang - - 2005
AIM: Transforming growth interacting factor (TGIF) is an inhibitor of both transforming growth factor beta (TGF-beta) and retinoid signaling pathways. Moreover, the activation of MAPK pathway can prolong its half-life. However, its role in carcinogenesis is still unknown. Thus we attempted to investigate the effect of TGIF on biologic behaviors ...
Derouazi, Madiha
Recombinant proteins are gaining in importance for therapeutic applications. The proteins are expressed in stable cell lines, within which recombinant DNA has incorporated into the host cells genome. Identification and isolation of extremely high producers from transfected cell populations is a tedious and time-consuming effort. We proposed two different approaches ...
Muller, Natalie
Recombinant proteins are important for biomedical research and for the treatment of human disease. Therefore it is necessary to develop reproducible bioprocesses to rapidly produce proteins of adequate quality and quantity. Expression in mammalian cells is preferred if the proteins are to be properly folded and post-translationally modified. For the ...
Iwamori Masao - - 2005
Transfection of the mouse Fut1 and Fut2, and human FUT1 genes into human ovarian carcinoma-derived RMG-1 cells resulted in 20-30-fold increases in cellular alpha1,2-fucosyltransferase activity, and in alteration of the glycolipid composition, including not only fucosylated products, but also precursor glycolipids. Although globo-series glycolipids were not significantly affected by the ...
Zheng Rui - - 2005
CD9, a transmembrane protein known as motility-related protein-1, plays a pivotal role in regulating cell adhesion, motility, and proliferation, and has been regarded as an important metastasis-inhibitory factor of various human cancers. However, little information has been obtained regarding the highly metastatic human small-cell lung cancer (SCLC). In the present ...
Tang Yi - - 2005
The transfection efficiency of oligonucleotide and plasmid to the HL-60 cell line with lipofectaminePLUS was compared through observing the transfection rate and the expression duration of exogenous gene in the target cells. The results showed that the transfection rate of oligonucleotide to the HL-60 was about 90% - 95% and ...
Liang Fong-Qi - - 2005
Glutathione-S-transferases (GSTs) play an important role in protection mechanisms against oxidative stress. We sought to determine whether over-expression of human GSTA1-1 in RPE cells is able to attenuate H(2)O(2)-induced oxidative stress. SV40-transformed human fetal RPE cells were stably transfected with pRC/hGSTA1-1 vector which carries a full-length of human GSTA1-1 cDNA. ...
Wu Yue - - 2005
Transient transfection of NIH3T3 cells with various constructs of myosin phosphatase target subunit (MYPT1) and GFP showed distinct cellular localizations. Constructs containing the N-terminal nuclear localization signals (NLS), i.e. full-length MYPT1 and N-terminal MYPT1 fragments, were concentrated in the nucleus. Full-length chicken and human MYPT1-GFP showed discrete nuclear foci. Deletion ...
Lu Xingrong - - 2005
In order to investigate the effect of small ubiquitin-like modifier-1 (SUMO-1) on the p53-induced HepG2 cell apoptosis, HepG2 cells were transfected by recombinant plasmids as pwtp53, pMDM2 and pSUMO-1 respectively. Western blot was employed to detect the protein expression of the transfected recombinant plasmids and the rate of apoptosis was ...
Liao Hongtao - - 2005
Small interfering RNAs (siRNA) could be the ideal inhibitor of specific gene expression if they could be delivered efficiently to their targets. Although siRNAs cannot by themselves cross cell membranes, they can enter cells with the help of transfection reagents via facilitated transport. Inside cells, however, they can still be ...
Adams Peter D - - 2005
Short interfering (si) RNAs are commonly used to knock down expression of proteins in mammalian cells and thereby investigate protein function. siRNAs were originally introduced into mammalian cells by transient transfection of short, synthetic, double-stranded RNA oligonucleotides. More recently, a convenient, more cost-effective approach has been developed that makes use ...
Topaloglu Ozlem - - 2005
Recent studies have demonstrated the utility of recombinant adeno-associated viral (rAAV) vectors in the generation of human knockout cell lines. The efficiency with which such cell lines can be generated using rAAV, in comparison with more extensively described plasmid-based approaches, has not been directly tested. In this report, we demonstrate ...
Wagner Ernst - - 2005
Dominating issues in gene vector optimization are specific in recognizing the target cells and exploiting the proper intracellular trafficking routes. Any progress in this area will result in improved specific gene transfer, reduce the required therapeutic vector doses and, in consequence, lower the overall toxicity to the host. To provide ...
Reed Chantal A - - 2005
In an effort to optimize the transfection of cell lines with antisense oligonucleotides, we examined cellular accumulation of a labeled oligonucleotide by flow cytometry. We were surprised to observe that a routinely used transfection protocol, a fixed lipid/oligonucleotide ratio, resulted in variable transfection efficiency depending on the concentration of oligonucleotide ...
Borth Nicole - - 2005
Previous work has shown that a human-antibody-producing recombinant CHO cell line did not increase its intracellular content of protein disulfide isomerase (PDI) and heavy chain binding protein (BIP) according to the increasing expression of antibody. It was also found that the intracellular assembly of light and heavy chain is a ...
Wang Li - - 2005
As previously shown, vinblastine, when incorporated into a cationic lipid prior to generation of lipoplexes, increases by approximately 30-fold the extent of transfection of pbeta-Gal with a cytomegalovirus promoter (pCMV-beta-Gal) to vascular smooth muscle cells (VSMC) by 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (EDOPC)-pCMV-beta-Gal complexes. To test if this increase is limited to VSMC and ...
Clement Jule - - 2005
The instability of lipoplex formulations is a major obstacle to overcome before their commercial application in gene therapy. In this study, a continuous mixing technique for the large-scale preparation of lipoplexes followed by lyophilisation for increased stability and shelf-life has been developed. Lipoplexes were analysed for transfection efficiency and cytotoxicity ...
Laitala-Leinonen Tiina - - 2005
Bone-resorbing osteoclasts are multinucleated cells that are formed via fusion of their hematopoietic stem cells. Many of the details of osteoclast formation, activation and motility remain unsolved. Therefore, there is an interest among bone biologists to transfect the terminally differentiated osteoclasts and follow their responses to the transgenes in vitro. ...
Chen Yong Y Department of Chemical and Biological Engineering, Northwestern University, Evanston, Illinois 60208, - - 2005
Pantropic retroviral vectors pseudotyped with vesicular stomatitis virus envelope G protein (VSV-G) are typically produced by transient transfection of the VSV-G expression plasmid because constitutive expression of VSV-G is cytotoxic. To produce pantropic vectors, the VSV-G expression plasmid and the vector plasmid are cotransfected into a packaging cell line, such ...
Berrier Allison L AL LSU School of Dentistry, Department of Cell Biology and Anatomy, New Orleans, LA, - - 2005
A method is provided to quantitate the extent of cell spreading as a function of the expression level of transfected recombinant proteins. This chapter contains protocols for 1) replating and staining transfected cells for immunofluorescence microscopy, 2) optimizing image acquisition so that fluorescence intensity can be measured independent of cell ...
Tait Andrew S - - 2004
We have developed a simple and robust transient expression system utilizing the 25 kDa branched cationic polymer polyethylenimine (PEI) as a vehicle to deliver plasmid DNA into suspension-adapted Chinese hamster ovary cells synchronized in G2/M phase of the cell cycle by anti-mitotic microtubule disrupting agents. The PEI-mediated transfection process was ...
Wang Xiaolin - - 2004
Tissue factor (TF) is the membrane receptor of the serine protease coagulation factor VIIa (FVIIa). Formation of the TF/FVIIa complex initiates the coagulation cascade. We used short hairpin RNA (shRNA)-mediated RNA interference to knock down TF expression in the human metastatic melanoma cell line LOX-L. After transfection with the shRNA ...
Hung Chi-Feng - - 2005
Transfection systems based on complexes of DNA and lipid emulsions were evaluated with respect to their effectiveness, toxicity, physicochemical characteristics, and cell-type dependence. The potential of a series of co-emulsifiers to serve as vectors was investigated. The co-emulsifiers examined included 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), Tween, cholesterol, stearylamine, and polyethylenimine (PEI). Squalane and ...
Delehanty James B - - 2004
The expression of recombinant antibody fragments on the surface of mammalian cells has recently emerged as a therapeutic strategy, particularly in the treatment of a number of cancers. Screening technologies that allow for the facile characterization of fragments expressed on the cell surface would hasten the identification and isolation of ...
Xu Shou-Shui - - 2004
AIM: To study the inhibitory effect of transfected PTEN on LoVo cells. METHODS: Human PTEN cDNA was transferred into LoVo cells via lipofectin and PTEN mRNA levels and its expression were analyzed by Western blot and flow cytometry. Before or after transfection, the effects of 5-Fu on inhibiting cell proliferation ...
Dailey L A - - 2004
This study examined the effect of nebulization on the cellular uptake and transfection efficiency of polyplexes from four polyethylenimine (PEI) modifications: branched 25 kDa PEI (bPEI), linear 22 kDa PEI (linPEI), pegylated PEI (pegPEI) and biodegradable PEI (bioPEI). Polyplexes were aerosolized with air-jet and ultrasonic nebulizers. The aerosol was collected ...
Farson Deborah - - 2004
BACKGROUND: One of the major limitations to the use of adeno-associated virus (AAV) vectors for gene therapy has been the difficulty in producing enough vector to supply a clinical trial. More than 20 000 roller bottles may be required to generate AAV by the traditional transient transfection process to treat ...
Jin Ying - - 2004
1. This study describes the use of an oligodendroglial cell line (158N) to study the protective effects of X-chromosome-linked inhibitor of apoptosis (XIAP) overexpression. 2. 158N cells were transiently transfected with either pCMV-Myc-XIAP or control pCMV-Myc vector. At 48 h post-transfection, immunoblotting and immunocytochemical staining showed robust myc-XIAP overexpression in ...
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