Navigation Links
Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN ,,, siRNA and TransMessenger Transfection Reagent*

Anna M. Krichevsky and Kenneth S. Kosik
Center for Neurologic Diseases, Brigham and Womens Hospital, Harvard Medical School, Boston, MA, USA Genetic manipulations using neuronal cells are notoriously difficult. Therefore, efficient delivery of short interfering RNA (siRNA) to primary neuronal cultures is of critical importance for RNA interference (RNAi)-mediated gene suppression. Here we describe successful targeted suppression of gene expression using TransMessenger Transfection Reagent and synthetic siRNA directed against microtubule-associated protein 2 (MAP2). RNAi has become a powerful tool used to knock down the expression of genes in cultured mammalian cells. However, neuronal cells, which are difficult to manipulate, have seemed to be more resistant to RNAi than other cell types. The reasons for this are unclear. Differences in siRNA uptake across the neuronal cell membrane or the RNAi pathway itself are possible causes. In this study, synthetic siRNA was used to successfully suppress expression of MAP2.

Materials and methods Two QIAGEN siRNAs targeted against MAP2 mRNA were designed with a 5' phosphate, 3' hydroxyl, and two-base overhangs on each strand (see Table "siRNA sequences used to target endogenous MAP2 mRNA in neuronal cells"). Transfections of siRNA for gene targeting were performed using TransMessenger Transfection Reagent as described in the TransMessenger
'"/>


Source:


Page: All 1 2 3 4 5

Related biology technology :

1. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
2. Efficient Cleavage of Fusion Proteins to Yield Native Amino Termini
3. Mammalian Expression Vector for Efficient Cloning of PCR Fragments
4. Efficient Transfection of Neurospora Crassa
5. Efficient Transfection of Neurospora Crassa
6. Efficient Recovery of Ultrapure Plasmid DNA
7. Efficient and Reliable PCR Setup Using Eppendorf MasterMix
8. Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent
9. Cloning Based on Efficient Three-Fragment Assembly DNA Ligation
10. Efficient and Reliable Linear Amplification of cRNA
11. Efficient Delivery of siRNAs to Human Primary Cells: Electroporation vs. Chemical Transfection
Post Your Comments:
(Date:7/28/2014)... July 28, 2014 Research and Markets ... Synthetic Biology Market 2014-2018" report to their offering. ... The term synthetic biology covers the designing and engineering ... well as the redesigning of natural biological systems that ... be a form of extreme genetic engineering because it ...
(Date:7/28/2014)... 28, 2014 According ... Market by Products (Consumables, Instruments, Services), Methods (Phenotypic, Genotypic), Applications (Pharmaceutical, ... (Detection, Characterization) - Global Forecasts to 2019" ... is estimated at $896.5 Million in 2014 ... by 2019, growing at a CAGR of ...
(Date:7/28/2014)... Boston, MA (PRWEB) July 28, 2014 ... with two researchers to further develop potential treatments for ... brain tumor, in combination with their patented vaccine platform, ... and Duke University, are based on the work of ... of Neurosurgery at The Preston Robert Tisch Brain Tumor ...
(Date:7/28/2014)... St Louis, MO (PRWEB) July 28, 2014 ... tools, software, and services that bring the power of ... for the inaugural Appistry Pipeline Challenge . ... Callum Bell, president of the National Center for Genome ... Children’s Mercy Hospitals and Clinics of Kansas City. Rounding ...
Breaking Biology Technology:Global Synthetic Biology Market 2014-2018 2Microbial Identification Market worth $1,194.1 Million by 2019 2Microbial Identification Market worth $1,194.1 Million by 2019 3String of Collaborations Launch Immunomic Therapeutics’ Brain Tumor Program Pipeline 2String of Collaborations Launch Immunomic Therapeutics’ Brain Tumor Program Pipeline 3String of Collaborations Launch Immunomic Therapeutics’ Brain Tumor Program Pipeline 4Appistry Names Industry Experts from the National Center for Genome Resources and Mercy Children's, Kansas City, to Judge Pipeline Challenge Competition 2
... 7, 2011 Reportlinker.com announces that a new ... Industrial Biotechnology China News 1103 ... The Chinese central government was reported to continue ... the Twelfth Five-Year (2011-2015) Plan. This act is ...
... and industrial research scientists and engineers, as well ... encounter a steep learning curve when developing common ... book just published by Springer promises to be ... The new MEMS Materials and Processes Handbook is ...
... The U.S. Food and Drug Administration today approved ... tool to treat brain aneurysms without performing open surgery. ... An aneurysm is a weak, enlarged area in an ... blood pressure can cause it to rupture, which can ...
Cached Biology Technology:Reportlinker Adds Industrial Biotechnology China News 1103 2Reportlinker Adds Industrial Biotechnology China News 1103 3MEMS Materials and Processes Handbook -- a comprehensive, practical resource 2FDA Approves New Device to Treat Brain Aneurysms 2FDA Approves New Device to Treat Brain Aneurysms 3
(Date:7/28/2014)... advance the genetic engineering of "fuel" crops for ... by researchers with the U.S. Department of Energy ... led by Lawrence Berkeley National Laboratory (Berkeley Lab). ... enables scientists to identify and characterize the function ... biosynthesis of plant cell walls. , "Our unique ...
(Date:7/28/2014)... virus killers to prevent and treat HIV infection has ... response these molecules can arouse as they get rid ... at the Weizmann Institute and the National Institutes of ... these molecules interferons around the time of ... the disease. Their research appeared in Nature . ...
(Date:7/28/2014)... Institute for Biologically Inspired Engineering at Harvard University ... be commercialized by a newly formed private company ... personalized medicine products. The announcement follows ... Technology Development (OTD) and the start-up Emulate Inc., ... human Organs-on-Chips platform. , "This is a ...
Breaking Biology News(10 mins):How sweet it is 2How sweet it is 3Interfering with interferon 2Wyss Institute's technology translation engine launches 'Organs-on-Chips' company 2Wyss Institute's technology translation engine launches 'Organs-on-Chips' company 3
... light, Stacie Grassano pulls out a tube. "This is a great ... Inside, a tree branch is speckled with white fluff. "It's growing ... brings the branch close to his eye. "Yes," he says, with ... For some, a fungus success story means nothing is growing at ...
... reveal in the 29 March 2007 issue of the ... important structures that control how genes are packaged and ... time, we are seeing in very high resolution on ... an organism's genes," said B. Franklin Pugh, professor of ...
... into a box glowing with fluorescent light, Stacie Grassano pulls out ... clear plastic up to her face. Inside, a tree branch is ... it to Scott Costa. , Costa brings the branch close to ... a fungus success story." , For some, a fungus success ...
Cached Biology News:Immunization rates hit record high in poor countries 2Immunization rates hit record high in poor countries 3Scientists reveal structure of gateways to gene control 2Scientists reveal structure of gateways to gene control 3Scientists reveal structure of gateways to gene control 4Fungal factories may save hemlock forests 2Fungal factories may save hemlock forests 3
Mouse monoclonal antibody raised against a partial recombinant STAU2. NCBI Entrez Gene ID = STAU2...
...
Mouse polyclonal antibody to NR2F2 - nuclear receptor subfamily 2, group F, member 2...
... Hybridization Wash Pack provides optimized blocking ... Preparation Quality Clarifier: Ready-to-use wash solutions ... SigmaScreen(tm) APS Coated Slides for Microarrays ... Hybridization Blocking Buffer: 1L • Microarray ...
Biology Products: