Abstract:
:RNA interference methodology suppresses specific gene expression, thus mimicking loss-of-function mutation and enabling in vitro and in vivo gene function analysis. Lipofectamine RNAiMAX, a new transfection reagent, has been confirmed high efficiency in delivering small interfering RNA (siRNA) into mesenchymal stem cells and neural stem cells. In this study, we used three transfection reagents (Lipofectamine RNAiMAX, Oligofectamine and Lipofectamine 2000) to deliver siRNA into human embryonic stem (hES) cells and compared the silencing efficiency of enhanced green fluorescent protein transgene and Oct4, a key regulator of pluripotency. As a result, siRNA can be delivered into hES cells more efficiently by Lipofectamine RNAiMAX compared with the other two transfection reagents and high efficient knockdown of target genes was obtained by Lipofectamine RNAiMAX even at a low concentration of siRNA. Quantitative real-time PCR showed approximately 90% knockdown of Oct4 transcript with cognate Oct4 siRNA transfection by Lipofectamine RNAiMAX compared to control siRNA. These results demonstrated that Lipofectamine RNAiMAX is an efficient and excellent transfection reagent for delivering siRNA into hES cells.
journal_name
Mol Biotechnoljournal_title
Molecular biotechnologyauthors
Zhao M,Yang H,Jiang X,Zhou W,Zhu B,Zeng Y,Yao K,Ren Cdoi
10.1007/s12033-008-9043-xsubject
Has Abstractpub_date
2008-09-01 00:00:00pages
19-26issue
1eissn
1073-6085issn
1559-0305journal_volume
40pub_type
杂志文章abstract::We report on a unified rapid betaine-based-PCR protocol for amplification of the (CAG)n region in Huntington disease (HD) and the (CGG)n region in Fragile X syndrome (FXS), followed by an electrophoretic separation on automated sequencer for precise determination of the triplet numbers. The high betaine concentration ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-010-9260-y
更新日期:2010-06-01 00:00:00
abstract::Over the last decade transgenic mouse models have become a common experimental tool for unraveling gene function. During this time there has been a growing expectation that transgenes resemble the in vivo state as much as possible. To this end, a preference away from heterologous promoters has emerged, and transgene c...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1385/MB:29:2:153
更新日期:2005-02-01 00:00:00
abstract::An alternative method to combine mutagenesis PCR with dITP and fragmentation by endonuclease V for directed evolution was developed. In comparison to the routine protocol for directed evolution, dITP was used as mutation reagent in the mutagenesis PCR. Subsequently, the incorporated dITP in the PCR products could repr...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9516-9
更新日期:2013-01-01 00:00:00
abstract::Dye-based protein determination assays are widely used to estimate protein concentration, however various reports suggest that the response is dependent on the composition and sequence of the protein, limiting confidence in the resulting concentration estimates. In this study a diverse set of model proteins representi...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-007-0038-9
更新日期:2007-10-01 00:00:00
abstract::Several mutations were introduced into the Cry1Ac toxin gene, resulting in four variants with altered sequences that were responsible for low expression of the toxin in transgenic plants. These variants were as follows: V1, with modified three A/T-rich regions, including the first signal of transcription termination; ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/mb:26:1:17
更新日期:2004-01-01 00:00:00
abstract::In this study, host-specific forms of the blast pathogen Magnaporthe oryzae in sub-Saharan Africa (SSA) were characterised from distinct cropping locations using a combination of molecular and biological assays. Finger millet blast populations in East Africa revealed a continuous genetic variation pattern and lack of ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-011-9429-z
更新日期:2012-02-01 00:00:00
abstract::Association studies using genome scans to identify quantitative trait loci for multifactorial disorders, with anything approaching reasonable power, have been compromised by the need for a very dense array of genetic markers and large numbers of affected individuals. These requirements impose enormous burdens on the g...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/mb:22:3:253
更新日期:2002-11-01 00:00:00
abstract::Human immunodeficiency virus types 1 and 2 (HIV-1, -2), the etiological agents of AIDS, are retroviruses that replicate in CD4+ T lymphocytes, monocytes, and macrophages. Early anti-HIV therapies were directed at the step in the virus life cycle that was considered the most readily blocked: the transcription of viral ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02821510
更新日期:1994-02-01 00:00:00
abstract::Despite the known importance of the composition of culture media and culture conditions on Bacillus thuringiensis growth and toxicity, very few reviews are concerned with this subject. This article reviews some aspects of the microbiology of Bacillus thuringiensis, and how toxicity is affected by the composition of gr...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1007/BF02907471
更新日期:1995-08-01 00:00:00
abstract::We have modified the differential display of 3'-end restriction fragments of cDNA technique by combining it with the amplified fragment length polymorphism (AFLP) approach and silver staining. Modified oligo-dT primers were used for a reverse transcription step. ds cDNA was digested with the Mse I restriction enzyme a...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:12:2:137
更新日期:1999-09-01 00:00:00
abstract::A total of 41 accessions of Aglianico belonging to three different biotypes (Taburno, Taurasi, and Vulture) and 9 accessions of Sirica grapes were sampled from diverse areas of Campania (Italy). All accessions were first genotyped using 21 microsatellite markers (SSR) to evaluate possible homonymies, synonymies, and t...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9605-9
更新日期:2013-06-01 00:00:00
abstract::The induction of several secondary metabolites in plants is one of the most commonly observed effects after the external addition of methyl jasmonate (MeJA). After the elicitation of Catharanthus roseus hairy roots with different concentrations of MeJA, changes in the accumulation of alkaloids such as ajmalicine, serp...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-008-9111-2
更新日期:2009-03-01 00:00:00
abstract::We investigated the efficiency of a single plasmid transfection along the longitudinal axis of the regenerating soleus of young rats. This also reflected transfection efficiency along the fibers because the soleus is a nearly fusiform muscle in young animals. The complete regeneration was induced by notexin and the tr...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9555-2
更新日期:2013-06-01 00:00:00
abstract::Podophyllotoxin (PPT) is a plant natural product that serves as a precursor for the synthesis of many well-known chemotherapeutic drugs. The limited availability and high demand for the source plants of PPT have led to the exploration of alternative sources for this compound. In this study, we utilized the endophytic ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-015-9888-8
更新日期:2015-12-01 00:00:00
abstract::In an effort to develop microalgae as a robust system for the production of valuable proteins, we analyzed some of the factors affecting recombinant protein expression in the chloroplast of the green alga Chlamydomonas reinhardtii. We monitored mRNA accumulation, protein synthesis, and protein turnover for three codon...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-010-9348-4
更新日期:2011-05-01 00:00:00
abstract::High-quality RNA preparations are critical for further applications such as reverse transcriptase-polymerase chain reaction (RT-PCR) transcript amplifications, and elaboration of cDNA and expressed sequence tag libraries. Melanins are phenolic compounds present in many fungi and apparently play key roles in fungi path...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:34:1:45
更新日期:2006-09-01 00:00:00
abstract::PREP1/p160 is a protein complex with relevant physiopathological roles in vivo. p160 regulates PREP1 transcriptional activity by preventing the formation of other PREP1-containing complexes, whereas PREP1 regulates p160 activity by increasing its stability. This induces the repression of the insulin-regulated glucose ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-016-9932-3
更新日期:2016-05-01 00:00:00
abstract::Lentivirus-mediated RNA interference (RNAi) is a potent experimental tool for investigating gene functions in vitro and in vivo. It has advantages that transgenic technology lacks. However, in vivo applications are difficult to apply in the central nervous system of non-model organisms due to the lack of a standard br...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-012-9596-6
更新日期:2013-06-01 00:00:00
abstract::The ability to identify disease-associated genes using microarray technology is dependent on isolating high-quality total RNA from the diseased tissue under study. However, it is not always possible to obtain large amounts of affected tissue from patients and therefore the quality and yield of RNA may be compromised. ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:23:2:171
更新日期:2003-02-01 00:00:00
abstract::In this study, we investigated expression and dimerization of an ER-associated degradation (ERAD) substrate, a null Hong Kong variant of α-1-antitrypsin (NHK) using immunoblotting assay and a novel NanoLuc complementary reporter system called the NanoBiT (NB) assay. This NB-tagged NHK made it possible to monitor the i...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-018-0092-5
更新日期:2018-08-01 00:00:00
abstract::The biosensor field has grown enormously since the first demonstration of the biosensor concept by Leland C. Clark, Jr. in 1962. Today's biosensor market is dominated by glucose biosensors, mass-produced enzyme electrodes for the rapid self-diagnosis of blood glucose levels by diabetes sufferers. Here we take a histor...
journal_title:Molecular biotechnology
pub_type: 杂志文章,评审
doi:10.1385/MB:32:3:249
更新日期:2006-03-01 00:00:00
abstract::The filamentous fungus Sclerotinia sclerotiorum produces a complete set of cellulolytic enzymes needed for efficient solubilization of native cellulose, the major component of plants. In this work, we reported the molecular characterization of an important glycosyl-hydrolase enzyme classified as endo-β-1,4-glucanase. ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-013-9714-0
更新日期:2014-04-01 00:00:00
abstract::We describe an expression system for high-yield production of recombinant soluble human FasL (rsh- FasL) in CHO cells. After one round of selection for gene amplification, cell lines producing rsh-FasL up to 60 microg/L x 10(6) cells in 24 h were obtained. Cell lines were grown in protein-free medium as suspension cul...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:23:3:189
更新日期:2003-03-01 00:00:00
abstract::The simple use of nonisotopic hybridization probes to detect complementary sequences provides valuable information in a large number of research and commercial applications. In hybridization assays, the four "S's (speed, simplicity, sensitivity, and specificity) are important criteria for determining the choice of pro...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02740771
更新日期:1996-10-01 00:00:00
abstract::A functional assay for proteolytic processing of the amyloid precursor protein (APP) was set up in yeast. This consisted of a membrane-bound chimeric protein containing the beta-secretase cleaved C-terminal fragment of APP fused to the Ga14 transcription factor. Using this chimera in a GAL-reporter yeast strain, an ex...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:15:1:29
更新日期:2000-05-01 00:00:00
abstract::The aim of this study was to evaluate the usefulness of replicating but non disseminating adenovirus vectors (AdVs) as vaccine vector using human rotavirus (HRV) as a model pathogen. HRV VP7, VP4, or VP4Δ (N-terminal 336 amino acids of VP4) structural proteins as well as the VP4Δ::VP7 chimeric fusion protein were expr...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-013-9653-9
更新日期:2013-07-01 00:00:00
abstract::A duplex real-time PCR assay with melting curve analysis, using the EvaGreen fluorescence dye, was developed for rapid and reliable identification of bovine and caprine in ruminant feeds. The method merges the use of bovine (Bos taurus) and caprine (Capra hircus) specific primers that amplify small fragments (bovine 9...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/s12033-014-9756-y
更新日期:2014-08-01 00:00:00
abstract::A detailed method is presented for the creation of head-to-tail multimers of short blunt restriction fragments, ligated into a plasmid vector in a single-tube reaction. Random priming of the concatemer insert readily yields hybridization probes of high specificity, unattainable from the short monomer fragments. ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1007/BF02789291
更新日期:1994-08-01 00:00:00
abstract::Agrobacterium-mediated transformation is the method of choice to engineer desirable genes into plants. Here we describe a protocol for demonstrating T-DNA transfer from Agrobacterium into the economically important graminaceous plant maize. Expression of the T-DNA-located GUS gene was observed with high efficiency on ...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:13:2:165
更新日期:1999-12-01 00:00:00
abstract::Cyclooxygenase (COX)-2 plays an important role in the development of cancer and has been recognized as a potential therapeutic target. Because nonsteroidal anti-inflammatory drugs (NSAIDs) are able to inhibit the activity of this enzyme, the potential efficacy of such drugs for purposes of cancer prevention or therapy...
journal_title:Molecular biotechnology
pub_type: 杂志文章
doi:10.1385/MB:34:3:329
更新日期:2006-11-01 00:00:00