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Xu Fei - - 2010
Interleukin-22 is a novel cytokine produced mainly in activated T cells. The elaborate biological functions of IL-22 in vivo are still widely unknown. In this report, we describe a rapid, simple, and reproducible in vitro cell-based bioassay for measuring the bioactivity of recombinant interleukin-22 (IL-22) to study the primary function ...
Sommer Cesar A - - 2010
The residual presence of integrated transgenes following the derivation of induced pluripotent stem (iPS) cells is highly undesirable. Here we demonstrate efficient derivation of iPS cells free of exogenous reprogramming transgenes using an excisable polycistronic lentiviral vector. A novel version of this vector containing a reporter fluorochrome allows direct visualization ...
Stroh Thorsten - - 2010
The type of a nucleic acid and the type of the cell to be transfected generally affect the efficiency of electroporation, the versatile method of choice for gene regulation studies or for recombinant protein expression. We here present a combined square pulse electroporation strategy to reproducibly and efficiently transfect eukaryotic ...
Si-Tayeb Karim - - 2010
The use of lentiviruses to reprogram human somatic cells into induced pluripotent stem (iPS) cells could limit their therapeutic usefulness due to the integration of viral DNA sequences into the genome of the recipient cell. Recent work has demonstrated that human iPS cells can be generated using episomal plasmids, excisable ...
Berlivet Soizik - - 2010
RNA interference is widely used for loss-of-function studies in mammalian cells. As an alternative to the transfection of small RNAs, plasmid vectors have been developed to express short hairpin RNAs (shRNAs). We engineered the pHYPER shRNA vector, which is based on a 2.5-kb mouse genomic fragment encompassing the H1 gene. ...
Deng Yiping - - 2010
MMP-26 is a novel member of the MMP family and is widely expressed in cancer cells of epithelial origin. Published research shows that MMP-26 contributes to tumor development and to the restoration of tissue injury. In this study, in order to identify the functions of MMP-26 that contribute to the ...
Un Keita - - 2010
To achieve effective gene therapy, it is necessary to selectively and efficiently transfect therapeutic gene into targeted cells. In this study, we developed a combination method using mannosylated lipoplexes, which show selectivity to antigen-presenting cells such as macrophages and dendritic cells, and bubble liposomes (BLs), which are known to enhance ...
Moore Jennifer C - - 2010
Genetic manipulation of human embryonic stem cells (hESC) has been limited by their general resistance to common methods used to introduce exogenous DNA or RNA. Efficient and high throughput transfection of nucleic acids into hESC would be a valuable experimental tool to manipulate these cells for research and clinical applications. ...
Su Ying - - 2010
PURPOSE: Improper proliferation of lens epithelial cells is causally related to posterior capsule opacification. In the present study, we investigated whether small interfering RNA (siRNA)-mediated gene silencing of S-phase kinase-interacting protein 2 (Skp2) can be employed to inhibit rabbit lens epithelial cell (rLEC) proliferation by increasing the p27(kip1) level. METHODS: ...
Zhang Siyang - - 2010
Aberrant regulation in the invasion of cancer cells is closely associated with their metastatic potentials. TrkB functions as a receptor tyrosine kinase and is considered to facilitate tumor metastasis. Pyk2 is a non-receptor tyrosine kinase and integrates signals in cell invasion. However, little is known about the expression of TrkB ...
Li Shujun - - 2010
This study aimed to investigate the mechanism by which the human lung cancer drug resistance-related gene BC006151 regulates chemosensitivity by down-regulating BC006151 expression via antisense gene transfer in H446/(C)DDP cells. A retroviral vector containing the antisense BC006151 sequence was constructed and transfected into H446/(C)DDP cells. Transfection of the empty vector ...
Fjeldbo Christina Saeten - - 2010
RNA-transfected cell microarray shows great promise in functional genomics. By printing siRNA complexed with transfection reagent on glass slides, arrays of transfected cells are formed in which the phenotypic consequences of gene suppression can be investigated. Using reporter plasmids with fluorescence intensity as output data, we have developed a strategy ...
Huang Ying - - 2010
Wnt-5a is important in the physiological development and differentiation of lung and is also involved in the regulating proliferation, differentiation and invasion of tumor cells. However, very little is known about the roles of Wnt-5a in the development of lung cancer. The purpose of this study was to explore the ...
Hagemann Carsten - - 2010
Glioblastoma multiforme (GBM) are the most common malignant brain tumours in adults, characterized by short survival periods of patients. Their aggressive local growth pattern and increased invasiveness, due to a high motility of the tumour cells, hamper treatment. However, the molecular mechanisms regulating glioblastoma cell migration are still elusive. Here, ...
Choi Hye-Sun - - 2010
Since hepatocellular carcinoma (HCC) is one of the leading causes of cancer death worldwide, it is still important to understand hepatocarcinogenesis mechanisms and identify effective markers for tumor progression to improve prognosis. Amplification and overexpression of Tropomyosin3 (TPM3) are frequently observed in HCC, but its biological meanings have not been ...
Burger Katharina - - 2010
The Host Cell Reactivation Assay (HCRA) is widely used to identify circumstances and substances affecting the repair capacity of cells, however, it is restricted by the transfection procedure used and the sensitivity of the detection method. Primary skin cells are particularly difficult to transfect, and therefore sensitive methods are needed ...
Kang Sang-Gyun - - 2010
Prion diseases, which are called transmissible spongiform encephalopathies (TSEs), comprise a group of fatal infectious neurodegenerative disorders. Investigation of prion strains and generation of species dependent TSE model are necessary to understand pathogenesis of the disease. To establish a BSE-specific in vitro cell culture model, N2a and GT1 mouse neuronal ...
Liebig Tanja M - - 2010
Research on B cells has shown that CD40 activation improves their antigen presentation capacity. When stimulated with interleukin-4 and CD40 ligand (CD40L), human B cells can be expanded without difficulties from small amounts of peripheral blood within 14 days to very large amounts of highly-pure CD40-B cells (>10(9) cells per ...
He Yunfeng - - 2010
The hepaCAM gene encodes a new immunoglobulin-like cell adhesion molecule, and its expression is suppressed in a variety of human cancers. Additionally, hepaCAM possesses properties often observed in tumor suppressor genes. However, the expression and biological function of hepaCAM has not been investigated in bladder cancer. Therefore we sought to ...
Ho Ivy A - - 2010
Glioblastoma multiforme is the most malignant cancer of the brain and is notoriously difficult to treat due to the highly proliferative and infiltrative nature of the cells. Herein, we explored the combination treatment of pre-established human glioma xenograft using multiple therapeutic genes whereby the gene expression is regulated by both ...
Zhan Yaoyao - - 2010
The pair-rule gene, hairy, encodes a basic helix-loop-helix transcription factor and is required for patterning of the early Drosophila embryo and for morphogenesis of the embryonic salivary gland. Although hairy was shown to be expressed in the tracheal primordia and in surrounding mesoderm, whether hairy plays a role in tracheal ...
Wells Julie - - 2010
Compared to primary keratinocytes, HaCaT cells are easier to transfect and yet still maintain at least some features of normal epidermal proliferation and differentiation. This chapter describes methods used in our laboratory to maintain HaCaT cells in an undifferentiated state and to use the siRNA technology to efficiently deplete a ...
Cochrane Dawn R - - 2010
We focus on unique roles of miR-200c in breast, ovarian, and endometrial cancers. Members of the miR-200 family target ZEB1, a transcription factor which represses E-cadherin and other genes involved in polarity. We demonstrate that the double negative feedback loop between miR-200c and ZEB1 is functional in some, but not ...
Zhang Zhen-Ya - - 2010
STIM1, Orai1 and TRPC1 are all reported to be important for store-operated Ca(2+) entry (SOCE) in diverse cells. However, there is no evidence for the functional interaction of the three proteins in SOCE in human liver cells. The objective of this study is to determine whether they are involved in ...
Huang Fu-Wei - - 2009
Poly(L-succinimide)-graft-polyethylenimines (PSPs) were prepared as non-viral vectors for gene transfection. Branched polyethylenimine (Mw= 800, PEI800) was grafted to poly(L-succinimide) (PSI) in a one-step reaction with no catalyst. Gel retardation assay showed that the mobility of PSP/pDNA complexes was completely retarded at the low N/P ratio of 0.42. In vitro transfection ...
Koga Takahisa - - 2010
Myocilin and optineurin are two genes linked to glaucoma, a major blinding disease characterized by progressive loss of retinal ganglion cells (RGCs) and their axons. To investigate the effects of force-expressed wild-type and mutant myocilin and optineurin on neurite outgrowth in neuronal cells, we transiently transfected cells with pEGFP-N1 (mock ...
Zhou Li - - 2010
BACKGROUND: The overexpression of LAPTM4B-35 in gallbladder carcinoma (GBC) and its clinicopathologic and prognostic significance have been previously shown. Thus, this gene may play a role in the growth of GBC cells. METHODS: The pcDNA3-AE containing the complete open reading frame of LAPTM4B (lysosome-associated protein transmembrane-4beta) and mock (pcDNA3) plasmids ...
Lavorini-Doyle Cassidy - - 2009
The use of cationic lipid-DNA complexes (lipoplexes) for gene therapy of oral squamous cell carcinoma may be limited by the resistance of some cell types to transfection. Using fluorescence microscopy, fluorometry and luciferase luminescence, the authors examined whether the variability arises from the intracellular fate of lipoplexes. In transfection-resistant cells, ...
Cheng Bin - - 2010
OBJECTIVES: This study aimed at examining the effects of hepatitis B viral X protein (HBx) on the biological features and the expression of DNA repair enzymes in non-tumour human hepatic LO2 cells in vitro. METHODS: The HBx gene was transfected into LO2 cells to establish stably HBx-expressing LO2/HBx cells. The ...
Li Xi - - 2010
The unique physical property of thermo-responsive polymer (TRP) has recently prompted its increasing applications in tissue engineering. On the other hand, TRP has not been exploited for potential applications in quantitative cell screening against external stimulations. In this study, TRP is applied as a model system for elucidating the effect ...
Ross Jason W - - 2010
Development of a transgenic porcine biomedical research model requires effective delivery of DNA into the donor cell followed by selection of genetically modified somatic cell lines to be used for nuclear transfer. The objective of the current study was 2-fold: (1) to compare the effectiveness of a single 1 ms ...
Zheng Li-Duan - - 2009
AIM: To develop short hairpin RNA (shRNA) against heparanase, and to determine its effects on heparanase expression and the malignant characteristics of gastric cancer cells. METHODS: Heparanase-specific shRNA was constructed and transferred into cultured the gastric cancer cell line SGC-7901. Stable subclonal cells were screened by G418 selection. Heparanase expression ...
Ladewig Katharina - - 2010
Although siRNAs have surpassed expectations in experiments to alter gene expression in vitro, the lack of an efficient in vivo delivery system still remains a challenge in siRNA therapeutics development and has been recognized as a major hurdle for clinical applications. In this paper we describe an inorganic nanoparticle-based delivery ...
Zheng Yu-ping - - 2009
AIM: To investigate the effect of lentivirus-mediated integrin-linked kinase (ILK) RNA interference (RNAi) on human retinal Müller cells transdifferentiation into contractile myofibroblasts. METHODS: A lentiviral vector expressing ILK-specific shRNA was constructed and introduced into cultured retinal Müller cells. Silencing of the ILK gene was identified by real time RT-PCR and ...
Gouveia Ricardo - - 2010
L1 is a cell adhesion molecule that is heavily glycosylated and is essential for normal development of the central nervous system. In this work, we compare the N-glycosylation of the L1 mutant that consists of immunoglobulin domains 5 and 6 (L1/Ig5-6), expressed in insect Spodoptera frugiperda Sf9 and Trichoplusia ni ...
Chen Charng-Jui - - 2010
CEACAM1-4S (carcinoembryonic antigen-related cell adhesion molecule 1) is a type I membrane protein with a short (12-amino acid) cytoplasmic tail. Wild type CEACAM1-4S-transfected MCF7 cells form glands with lumena when grown in 3D culture, while null mutations of two putative phosphorylation sites (T457A and S459A) in the cytoplasmic domain fail ...
Reslan Lina - - 2010
Sonoporation holds many promises in developing an efficient, reproducible and permanent gene delivery vector. In this study, we evaluated sonoporation as a method to transfect nucleic acids in suspension cells, including the human follicular lymphoma cell line RL and fresh human Chronic Lymphocytic Leukemia (CLL) cells. RL and CLL cells ...
Lin E-H - - 2010
Fusogenic membrane glycoproteins (FMGs) are viral envelope proteins, which bind surface receptors and induce fusion of the cell membrane. An FMG-transfected cell will fuse with neighbor cells, thus forming syncytia that die within 5 days. In this report, plasmids encoding for FMGs from Human Endogenous Retrovirus-W (HERV-W) was compared with ...
Sengupta Manjistha - - 2010
Low-copy-number plasmids, such as P1 and F, encode a type Ia partition system (P1par or Fsop) for active segregation of copies to daughter cells. Typical descriptions show a single central plasmid focus dividing and the products moving to the cell quarter regions, ensuring segregation. However, using improved optical and analytical ...
McCulloch D R - - 2010
Glucosamine has been previously shown to suppress cartilage aggrecan catabolism in explant cultures. We determined the effect of glucosamine on ADAMTS5 (a disintegrin-like and metalloprotease domain (reprolysin type) with thrombospondin type-1 motifs 5), a major aggrecanase in osteoarthritis, and investigated a potential mechanism underlying the observed effects. HEK293F and CHO-K1 ...
Esendagli Gunes - - 2009
Gene transfer into haematopoietic cells is a challenging approach. The extracellular matrix component fibronectin has been known to modulate the cell cycle dynamics, viability and differentiation of leukaemia cells. Thus, our aim was to investigate the influence of fibronectin substratum on the liposomal transfection of myeloid leukaemia cell lines. Liposomal ...
Yu H - - 2009
The first establishment of a homologous expression system in the host Ralstonia pickettii CGMCC1596 using the compatible broad-host-range plasmid pWB5 is described. When whole cells of the recombinant strain R. pickettii MMYY01 (CGMCC1596/pYY05) were used as the biocatalyst to transform DL-4-hydroxyphenylhydantoin (DL-HPH) to D-4-hydroxyphenylglycine (D-HPG), the conversion rate reached 94 ...
Herringson Thomas P - - 2009
BACKGROUND: The ability to deliver plasmid DNA (pDNA) to specific cells in vivo is crucial for achieving efficient targeted transfection with nonviral vectors. We previously used stealth liposomes containing the chelator lipid 3(nitrilotriacetic acid)-ditetradecylamine (NTA(3)-DTDA) to target delivery of antigen and cytokines to immune cells in vivo. In the present ...
Dos Santos Alexandra Souza - - 2009
The aim of this study was to achieve expression of recombinant rabies virus glycoprotein (rRVGP) in Drosophila S2 cells. For this, a cDNA coding for the selection hygromycin antibiotic and the cDNA encoding the RVGP protein under the control of the constitutive actin promoter (Ac) were cloned in an expression ...
Mao Zhen Biao - - 2009
OBJECTIVE: To investigate the expression of guanylyl cyclase C (GCC) in human gastric cancer (GC) tissues and assess the effect of GCC small interfering RNA (siRNA) on the proliferation and apoptosis of SGC-7901. METHODS: The expression of GCC in 30 specimens and three human GC cell lines (SGC-7901, AGS, NCI-N87) ...
Ghonaim Hassan M - - 2010
PURPOSE: To study the efficiency of novel synthetic N(1),N(12)-diacyl spermines on DNA and siRNA binding, and to compare their transfection efficiency with viability in cell lines. METHODS: Five long chain N(1),N(12)-diacyl lipopolyamines: N(1),N(12)-[didecanoyl, dimyristoyl, dimyristoleoyl, distearoyl and dioleoyl]-1,12-diamino-4,9-diazadodecane were synthesized from the naturally occurring polyamine spermine. Their abilities to condense ...
Wang Feng - - 2010
PURPOSE: Although antiproliferative drugs have been used to prevent scarring after filtration surgery in patients with glaucoma, there are complications associated with their use. In the present study, the authors investigated whether small interfering RNA (siRNA)-mediated gene silencing of Skp2 can be used to increase p27(kip1) level and inhibit cell ...
Zumbansen Markus - - 2010
Meaningful RNAi-based data for target gene identification are strongly dependent on the use of a biologically relevant cell type and efficient delivery of highly functional siRNA reagents into the selected cell type. Here we report the use of the Amaxa(R) Nucleofector(R) 96-well Shuttle(R) System for siRNA screening in primary cells. ...
Kennard Malcolm L - - 2009
In order to maximize recombinant protein expression in mammalian cells many factors need to be considered such as transfection method, vector construction, screening techniques and culture conditions. In addition, the host cell line can have a profound effect on the protein expression. However, auditioning or directly comparing host cell lines ...
Kayama Maki - - 2010
OBJECTIVE: It is theoretically possible to induce various cell types, including retinal neurons, from embryonic stem cells (ESCs). pax6 regulates early events in eye development, including the generation of retinal ganglion cells (RGCs). We previously reported the successful induction of corneal epithelial cells from ESCs transfected with the pax6 gene. ...
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