Abstract：
:RNA interference (RNAi) is a powerful tool for functional genetic studies in model organisms and mammalian cells. To facilitate rapid construction of gene knockdown constructs and RNAi libraries for known genes of mammalian cells, a new and simple strategy to produce small interfering RNA (siRNA) expression vectors with two opposing polymerase III promoters was developed. The design involved a one-step PCR amplification and single cloning procedure to construct a dual promoter siRNA expression vector. The forward primer is identical for all PCR reactions, only a single reverse primer that contains the siRNA targeting sequence has to be synthesized in the construction of each individual vector. This single primer design is cost-effective and it reduces the risk of sequence errors during synthesis of long oligos. Sense and antisense strands of siRNA duplexes were transcribed from the same template and this eliminated the need to synthesize long hairpin-forming oligonucleotides. Our study demonstrated that this vector design could mediate potent inhibition of expression of both exogenous and endogenous genes in mammalian cells.
journal_name
Eur J Cell Bioljournal_title
European journal of cell biologyauthors
Jian R,Peng T,Deng S,Jiang J,Hu F,An J,Cheng Xdoi
10.1016/j.ejcb.2005.10.009subject
Has Abstractpub_date
2006-05-01 00:00:00pages
433-40issue
5eissn
0171-9335issn
1618-1298pii
S0171-9335(05)00192-5journal_volume
85pub_type
杂志文章abstract：:It is generally believed that the induction of actin cytoskeleton rearrangements by extracellular stimuli results from the activation of guanine nucleotide exchange factors for the Rho GTPases. Here, we present evidence that the inactivation of RhoGAP (GTPase activating protein) activity is an equally effective means ...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1016/S0171-9335(99)80019-3
更新日期：1999-08-01 00:00:00
abstract：:Monoclonal antibodies were used to define cell surface antigens which are present on rat hepatocytes but are absent from hepatoma cells. One monoclonal antibody, referred to as Be 9.2, recognizes a major component of purified rat liver plasma membranes with a Mr of 110 000. This antigen (gp110) was not found in the tr...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1986-01-01 00:00:00
abstract：:The major families of proteoglycans in human arterial tissue have been localized and characterized by electron microscopy. After staining with the polycationic dye cuprolinic blue in the presence of a critical electrolyte concentration, three differently sized populations of proteoglycan-cuprolinic blue precipitates a...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1987-12-01 00:00:00
abstract：:Dictyosomes, Golgi vesicles, and plasma membranes were investigated after freeze-fracture in cells from growing root tips of cress (Lepidium sativum L.), that are distinguishable by different cellulose content of the cell wall, into (i) meristematic cells during early formation of the cell plate, (ii) statocytes of th...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1983-05-01 00:00:00
abstract：:A monoclonal antibody (ID5) raised against the synthetic tetradecapeptide corresponding to the C-terminal region of detyrosinated alpha-tubulin showed an unexpected cross-reactivity with beta-tubulin from pig brain tissue. The specificity and the minimal epitope requirements of ID5 were characterized by competitive en...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1016/s0171-9335(99)80003-x
更新日期：1999-01-01 00:00:00
abstract：:The transport of the chitin synthase III, Chs3p, to the plasma membrane is temporally and spatially regulated. Chs3p is delivered to the plasma membrane at the beginning of the cell cycle, forming chitin rings, and at the end of the cell cycle, forming the primary septum. During the rest of the cell cycle, it is maint...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1016/j.ejcb.2005.10.006
更新日期：2006-02-01 00:00:00
abstract：:Interleukin-1 beta (Il-1 beta) and interleukin-1 alpha (Il-1 alpha) were shown to act as motility factors for the human breast carcinoma cell lines SK-BR-3 and ZR-75-1 in vitro. Both cytokines induced transition from the stationary to the motile phenotype (spreading). Il-1 beta stimulated translocation, shape change a...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1992-12-01 00:00:00
abstract：:Leydig cells in developing fetal pig testis contained during the fetal regressive phase large accumulations of intermediate filaments. Before and after this period these filaments were arranged in a criss-cross fashion. In the pig as well as in the dog testis these filaments have been characterized as vimentin. Within...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1984-07-01 00:00:00
abstract：:Mechanical tension governs fibroblast proliferation and survival and the homeostasis of the extracellular matrix to adapt its resistance to the mechanical requirements of the organs. To consolidate this view, we analysed the effect of tension release on the expression of molecules involved in the architecture and stab...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1078/0171-9335-00181
更新日期：2001-07-01 00:00:00
abstract：:Herein we report that Gts1p fused with green-fluorescent protein (GFP) is localized in the cortical actin patch besides nuclei in yeast and the cortical Gts1p changed its position together with the patch depending on the cell-cycle phase, while nuclear Gts1p accumulated predominantly in the budding phase. Whereas Gts1...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1016/j.ejcb.2007.02.005
更新日期：2007-05-01 00:00:00
abstract：:We have examined the polarity of microtubules in Sertoli cells of the seminiferous epithelium during testicular development to test the hypothesis that microtubules change their polarity during tight junction formation. Microtubules in a number of polarized epithelial cells, including Sertoli cells, are oriented with ...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1994-12-01 00:00:00
abstract：:In order to investigate the role of microfilaments in the crawling movements of lymphoid cells, we have analyzed the effects of botulinum C2 toxin and of cytochalasin D (cytoD) on the actin cytoskeleton and on the motility of a BW5147 T-lymphoma-derived cell line. Actin was ADP-ribosylated by C2 toxin in the living ce...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1995-04-01 00:00:00
abstract：:Extraovarian estrogen production is localized mainly in stromal vascular cells from adipose tissue, which contain the aromatase activity necessary for the conversion of androgens to estrogens. Induction of aromatase activity in these cells by glucocorticoids occurs only in the presence of a serum factor, the propertie...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1994-02-01 00:00:00
abstract：:It was previously shown that cultured mouse peritoneal macrophages ingest anionic derivatives of horseradish peroxidase (HRP) and ferritin by fluid-phase endocytosis and accumulate them in lysosomes. Cationic derivatives were taken up by adsorptive endocytosis and transported to lysosomes but subsequently appeared als...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1983-07-01 00:00:00
abstract：:Expression of the Cdx1 homeobox gene in epithelial intestinal cells promotes cellular growth and differentiation. Cdx1and the Pancreatitis Associated Protein I (PAP I) are concomitantly expressed in the epithelial cells of the lower part of the intestinal crypts. Because Cdx1 is a transcription factor and PAP I, in ot...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1078/0171-9335-00148
更新日期：2001-02-01 00:00:00
abstract：:Cartilage matrix protein (CMP), a major noncollagenous component of certain types of hyaline cartilage, is synthesized by chondrocytes in a developmentally regulated manner. In this study, we monitored the accumulation of CMP in the developing chicken limb and sternum by immunostaining. In older embryos, the specific ...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1995-12-01 00:00:00
abstract：:Fucoidan is a sulfated polysaccharide extracted from brown seaweeds. It has anticoagulant and antithrombotic properties and inhibits, as well as heparin, vascular smooth muscle cell growth. In this study, we investigated, in the presence of serum and human recombinant growth factors, the effects of fucoidan and hepari...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1016/S0171-9335(98)80094-0
更新日期：1998-12-01 00:00:00
abstract：:Three-dimensional collagen lattice cultures of fibroblasts mimic the in vivo situation better than monolayer cultures. Here, skin fibroblasts from scleroderma patients and healthy controls were cultivated in collagen lattices, and the effects of recombinant human gamma-interferon (IFN-gamma) on these cultures investig...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1992-04-01 00:00:00
abstract：:Human cytomegalovirus (HCMV) infection causes a rapid, progressive disruption of the host cell cytoskeleton that correlates with actin depolymerization. Whole-mount (3D) electron microscopy was used to analyze the cytoskeleton of uninfected and HCMV-infected human lung fibroblast cells. Within 2 min of HCMV infection,...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1986-08-01 00:00:00
abstract：:Storage and release of proteins from granules forms the basis of cellular functions as diverse as cell mediated cytotoxicity, neuronal communication, activation of muscle fibres, and release of hormones or digestive enzymes from endocrine and exocrine glands, such as the pancreas. Serglycin is the major intracellular ...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1016/j.ejcb.2009.03.002
更新日期：2009-08-01 00:00:00
abstract：:It has been known for many years that trypanosomatids require exogenous essential growth factors in order to divide. Two surface domains are involved in starting nutrient endocytosis: the flagellar pocket and the cytostome. Although the flagellar pocket plays a fundamental role in the endocytic process occurring in se...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1078/0171-9335-00112
更新日期：2000-11-01 00:00:00
abstract：:The transcriptional status of cells can be deduced from the staining pattern of various nuclear markers such as the Cajal body, nucleolus and nuclear speckles. In this study we have used these markers to correlate transcriptional status with cell differentiation in the lens. As a closed system with no cell loss and wi...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1078/0171-9335-00275
更新日期：2002-10-01 00:00:00
abstract：:The weak bases chloroquine, primaquine, NH4Cl and the ionophore monensin exert similar but not identical effects on sorting, transport and processing of cathepsin D in several human cell lines (fibroblasts, HepG2 cells, U937, monocytes). The drugs inhibit the segregation of newly synthesized cathepsin D from the secre...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1987-06-01 00:00:00
abstract：:Podosomes and the immune synapse are integrin-mediated adhesive structures that share a common ring-like morphology. Both podosomes and immune synapses have a central core surrounded by a peripheral ring containing talin, vinculin and paxillin. Recent progress suggests significant parallels between the regulatory mech...
journal_title：European journal of cell biology
pub_type： 杂志文章,评审
doi：10.1016/j.ejcb.2008.01.011
更新日期：2008-09-01 00:00:00
abstract：:A fodrin-like protein purified from porcine thyroid cells and characterized by its properties identical to those of pig brain spectrin (F. Regnouf et al., Eur. J. Biochem. 153, 313-319 (1985)) has been localized by immunofluorescence and electron immunocytochemistry in porcine and rat thyroid. Fodrin-like polypeptides...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1990-08-01 00:00:00
abstract：:Endogenous carbohydrate-binding sites were studied during rat cerebellar development on sections of fixed tissue using synthetic tools, biotinylated neoglycoproteins, in conjunction with subsequent avidinperoxidase staining. Neoglycoproteins were constructed by chemically coupling the histochemically pivotal carbohydr...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1990-06-01 00:00:00
abstract：:The differences in the content of trace elements (Fe, Cu, Zn, Br, Rb and Sr) in phasic and tonic scallop adductor muscles have been revealed by using X-ray fluorescence analysis. The phasic (yellow) muscles have been found to be richer in Fe and Cu while more deficient in Zn and Br as compared to the tonic (white) mus...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1981-04-01 00:00:00
abstract：:When resting human blood platelets are stimulated with thrombin, 50 to 60% of the G-actin polymerizes to F-actin within 60 seconds. The increase in F-actin is correlated with a corresponding decrease in the complex of G-actin with T beta 4. Within 5 seconds after stimulation, nucleation sites for pyrene actin polymeri...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1993-08-01 00:00:00
abstract：:The large protein bodies of the storage parenchyma cells of mung bean (Vigna radiata) cotyledons contain vesicles measuring 0.2 to 2.0 mum in diameter. The vesicles contain ribosomes, ribosomes, membranous elements which may be derived from the endoplasmic reticulum and occasionally Golgi bodies and mitochondria. The ...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：
更新日期：1981-06-01 00:00:00
abstract：:In recent years, several antagonists of alpha(v)beta3 have been used to develop therapeutic approaches to the treatment of melanoma neoplasia. We studied the effects of anti-alpha(v)-integrin-blocking antibodies on attached M21 melanoma cells, the cellular distribution of alpha(v)-integrin and the molecular organizati...
journal_title：European journal of cell biology
pub_type： 杂志文章
doi：10.1078/0171-9335-00067
更新日期：2000-07-01 00:00:00
