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Delivering 27mer DsiRNAs to Mice DRGsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
I have been a proponent of using 27mer DsiRNAs (Dicer Substrate Small Interfering RNAs) with our i-Fect kits to deliver siRNA to the CNS for gene expression analysis. The potency of this platform was highlighted in my profile of Dr. Mark Behlke.It was further confirmed by in Studies conducted by Dr. Philippe Serrat and his team at University of Sherbrooke.Louis Doré-Savard, Geneviève Roussy, Marc-André Dansereau, Michael A Collingwood, Kim A Lennox, Scott D Rose, Nicolas Beaudet, Mark A Behlke and Philippe Sarret. Central Delivery of Dicer-substrate siRNA: A Direct Application for Pain Research. Molecular Therapy (2008)...
Source: siRNA and DsiRNA Transfection Efficiency - June 23, 2009 Category: Neuroscience Tags: siRNA delivery in-vivo Dr. Mark Behlke Delivering siRNA to the CNS Dicer Substrate intrathecal delivery of siRNA Source Type: organizations

Knockdown of rSNSR1 in vivoEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The parade of success with use our i-FectTM in vivo grows. Here's the most recent study:Christian Ndong, Amynah Pradhan, Carole Puma, Jean-Pierre Morello, Cyrla Hoffert, Thierry Groblewski , Dajan O’Donnell, Jennifer M.A. Laird. Role of rat sensory neuron-specific receptor (rSNSR1) in inflammatory pain: Contribution of TRPV1 to SNSR signaling in the pain pathway. PAIN 143 (2009) 130–137....For experiments in which siRNA was delivered by bolus injections, 10 ul of siRNA or vehicle was injected directly into the intrathecal catheter once daily for 4 days. In this case, siRNAs were prepared immediately prior to administra...
Source: siRNA and DsiRNA Transfection Efficiency - April 20, 2009 Category: Neuroscience Tags: Dr. Jennifer Laird snsr1 IT Delivery of siRNA in vivo RNAi rat sensory neuron-specific receptor TRPV1 i-Fect intrathecal delivery of siRNA Source Type: organizations

Did Your RNAi Experiment Work?Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This is a good methods publication on RNAi transfection: Reliably Validating RNA Interference with qRT-PCR.Bill Wang, Song Tian, Qiong Zhou, and Xiao Zeng. SA Biosciences.
Source: siRNA and DsiRNA Transfection Efficiency - April 6, 2009 Category: Neuroscience Tags: Nearly 100% Transfection Efficiency siRNA siRNA Delivery RNAi PCR Source Type: organizations

Intrathecal Delivery of siRNAEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We wanted to present yet another publication referencing successful delivery of siRNA using i-FectTM:Suneeta Tumati, Tally Largent Milnes, Henry I. Yamamura, Todd W. Vanderah, William R. Roeske and Eva V. Varga. Intrathecal Raf-1-selective siRNA attenuates sustained morphine-mediated thermal hyperalgesia. doi:10.1016/j.ejphar.2008.10.033...siRNAs stock solutions (100 μM) were prepared in double distilled RNAse free water and stored in aliquots at −80 °C. For intrathecal treatment, aliquots of the stock solution (2 μg of the appropriate siRNA) were mixed (1:5 v/v)with i-Fect transfection reagent (Neuromics, Edina, MN)....
Source: siRNA and DsiRNA Transfection Efficiency - December 16, 2008 Category: Neuroscience Tags: Dr.Eva Varga siRNA delivery in-vivo IT Delivery of siRNA in vivo RNAi i-Fect intrathecal delivery of siRNA Source Type: organizations

Delivering Naked siRNA by Direct InjectionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, they delivered chemically modified murine and human alpha-synuclein (SNCA) siRNAs to the hippocampus by direct injection resulting in silencing of gene expression.To learn more access:In vivo silencing of alpha-synuclein using naked siRNA Jada Lewis, Heather Melrose, David Bumcrot, Andrew Hope, Cynthia Zehr, Sarah Lincoln, Adam Braithwaite, Zhen He, Sina Ogholikhan, Kelly Hinkle, Caroline Kent, Ivanka Toudjarska, Klaus Charisse, Ravi Braich, Rajendra K. Pandey, Michael Heckman, Demetrius M Maraganore, Julia Crook, Matthew J Farrer. Molecular Neurodegeneration 2008, 3:19 (1 November 2008).
Source: siRNA and DsiRNA Transfection Efficiency - November 2, 2008 Category: Neuroscience Tags: Mayo Clinic siRNA delivery in-vivo siRNA direct injection SNCA Parkinson's Disease. Dr. Matthew Farrer naked siRNA Alnylam Therapeutics Alpha-synuclein Gene Silencing PD Source Type: organizations

ACIC3 Receptors Knockdown in vivoEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
In this study, Dr. Eric Lingueglia and his team found Peripheral ASIC3 channels are thus essential sensors of acidic pain and integrators of molecular signals produced during inflammation where they contribute to primary hyperalgesia.Emmanuel Deval, Jacques Noël, Nadège Lay, Abdelkrim Alloui, Sylvie Diochot, Valérie Friend, Martine Jodar, Michel Lazdunski and Eric Lingueglia. ASIC3, a sensor of acidic and primary inflammatory pain. The EMBO Journal advance online publication 16 October 2008; doi: 10.1038/emboj.2008.213Cy3-labelled siRNA no. 1121 and its corresponding scramble (no. 1121S; GCTCACACTACGCAGAGAT) synthesized...
Source: siRNA and DsiRNA Transfection Efficiency - October 19, 2008 Category: Neuroscience Tags: siRNA delivery in-vivo IT infection siRNA Complexes i-Fect Eric Lingueglia iFect Source Type: organizations

siRNA-mediated gene silencingEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Dr. Josephine Lai (Professor of Pharmacology, University of Arizona) is pioneer in developing experimental designs and methods for delivering siRNA to the CNS for gene expression analysis.She and her team have documented these in the publication:Modulating Sensory Systems Using RNAi(pdf - 187Kb)© 2007 LaiFor researchers desiring to effectively deliver siRNA to the CNS for gene expression analysis of specific receptors, this publication offers proven methods. These include:The Choice of siRNA Choosing and Optimizing Transfection Reagents for siRNA Delivery to the Nervous SystemDelivery Systems-Microinjection and Infusion (...
Source: siRNA and DsiRNA Transfection Efficiency - October 4, 2008 Category: Neuroscience Tags: siRNA delivery in-vivo siRNA Microinjection Delivering siRNA to the CNS RNAi Gene Silencing Dr. Josephine Lai Source Type: organizations

Transfecting Sympathetic NeuronsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
GDNF and Ret are important to the growth, maintenance and survival of Neurons. The GDNF ligands act via activation of Ret. Every step towards understanding the intricacies of this pathway, brings researchers closer towards unlocking the code for Neurodegenerative Disease therapies.siRNA is an important tool for studying the Neurotrophic pathways as researchers can use it to modulate the expression of related receptors. The tricky part is getting sufficient siRNA into neurons to do the appropriate studies of how modulating targeted genes results in changes in protein expression.Here Drs. Cynthia Tsui and Brian Pierchala hav...
Source: siRNA and DsiRNA Transfection Efficiency - August 28, 2008 Category: Neuroscience Tags: Dr. Brian Pierchala GDNF; Ret; sympathetic neuron; neurotrophic factor; degradation; E3-ligase Dr. Cynthia Tsui Transfecting Neurons i-Fect Source Type: organizations

Down Regulating the Smad and Neuro-regenerationEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The inventors down regulated Smad 2/3 (an inhibitor of neuro-regeneration) in vivo via delivery of siRNA to the spinal cord using catheters.Inhibiting smad signaling promotes neuron regeneration. Inventors: Fan Wang, Zhigang HeUSPTO Application #: 20080031911Inhibition of Smad2/3 Signaling Promotes Axonal Regeneration after Spinal Injury in Rats-Gene Expression Knockdown in vivo!One hour after the spinal cord is lesioned, the rats in the SB-505124 group receive a bolus injection of SB-505124 (30 mg/kg) in 0.9% saline administered via a tail vein. The treatment is repeated every 24 hours on days 1 through 7 post-lesion. Veh...
Source: siRNA and DsiRNA Transfection Efficiency - August 25, 2008 Category: Neuroscience Tags: neuro-regeneration smad2/3 tgf-beta i-Fect Source Type: organizations

siRNA and DiabetesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
I'll be keeping my eyes on this and post updates.Quark Pharmaceuticals Announces First Patient Dosing by Pfizer in Phase II Trial of RNAi Therapy in Diabetic MClinical Program Leverages Quark's RNAi Technology FREMONT, Calif., July 30 /PRNewswire/ -- Quark Pharmaceuticals, Inc., a development-stage pharmaceutical company discovering and developing novel RNA interference (RNAi)-based therapeutics,...
Source: siRNA and DsiRNA Transfection Efficiency - August 1, 2008 Category: Neuroscience Tags: siRNA Diabetes Source Type: organizations

Improving 27mer DsiRNA PerformanceEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
The DsiRNA story marches forward with yet another important publication. We will give you an opportunity to upload the fulltext article at the end of this posting. Dr. Mark Behlke, Dr John Rossi and team have been gaining deeper understanding of the Mechanism of Dicer-substrate small-interfering RNA (DsiRNA) processing. This understanding is leading to better and better designs of the RNA duplexes. These designs or chemical modifications are necessary steps in the drug design and development process. This publication looks at design from the perspective of: Nuclease StabilityPharmacokinecticsimmune response I believe this ...
Source: siRNA and DsiRNA Transfection Efficiency - July 21, 2008 Category: Neuroscience Tags: Dr. Mark Behlke dsRNA Design Dr. John Rossi Dicer Complex Dicer-RISC DsiRNA Processing Source Type: organizations

Coming SoonEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Agenda Topics:Target Discovery and ValidationTransfectionsiRNA Therapeuticsin vivo RNAi-recent in vivo RNAi PubsBioinformatics of small RNAssiRNA library screensmicroRNAs inDisease Biology:Stem Cell BiologyDiagnosticsVirologyBiogenesisDevelopment
Source: siRNA and DsiRNA Transfection Efficiency - July 8, 2008 Category: Neuroscience Tags: microRNAs in vivo RNAi siRNA Therapeutics Transfection Source Type: organizations

RNAi Researchers Galvanized by AdvancesEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Technology's Viability in Drug Development Is Finally EstablishedAuthor: Elizabeth LippPublication: Genetic Engineering & biotechnology NewsPublisher: Mary Ann Liebert, Inc. publishersDate: Jun 1, 2008Copyright © 2008 GEN PublishingArticle Link: http://www.genengnews.com/articles/chitem.aspx?aid=2493Notable Quotables:“Long dsRNAs have been employed for many years as a means to modulate gene expression in plants, yeast, and C. elegans,” noted Mark Behlke, M.D., Ph.D., svp of molecular genetics and CSO at Integrated DNA Technologies (IDT; www.idtdna.com).“Similar attempts in higher organisms failed due to interfer...
Source: siRNA and DsiRNA Transfection Efficiency - June 18, 2008 Category: Neuroscience Tags: Mark Behlke siRNA Dicer Complex RNAi idtdna dsiRNA Source Type: organizations

Hi Thru Put Delivery of siRNA/DsiRNAEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
NEW PRODUCTi-Fect ™ Hi Put 96-NewLearn about transfection products @ neuromics
Source: siRNA and DsiRNA Transfection Efficiency - June 16, 2008 Category: Neuroscience Tags: in vitro Delivery RNAi dsiRNA Source Type: organizations

Central Delivery of DsiRNAEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Louis Doré-Savard, Geneviève Roussy, Marc-André Dansereau, Michael A Collingwood, Kim A Lennox, Scott D Rose, Nicolas Beaudet, Mark A Behlke and Philippe Sarret. Central Delivery of Dicer-substrate siRNA: A Direct Application for Pain Research. Molecular Therapy (2008); doi:10.1038/mt.2008.98.Images: Cellular uptake of Texas Red–tagged Dicer-substrate small-interfering RNA (DsiRNA) by spinal nociceptive structures. (a,b) Distribution of fluorescence in lumbar dorsal root ganglia at 24 hours after intrathecal injection of a control siRNA conjugated with Texas Red (1 μg administered twice with a 24-hour interval; n = 3...
Source: siRNA and DsiRNA Transfection Efficiency - June 4, 2008 Category: Neuroscience Tags: Mark Behlke siRNA Complexes in vivo iFect dsiRNA Source Type: organizations

Ret Knockdown Using Small Interference RNAEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Nearly 100% Transfection Efficiency Reported in vitro with i-Fect ™.Images: Ret receptor knockdown using small interference RNA (siRNA) in podocytes. (A) Transfection efficiency: mouse podocytes were transfected with 100 nM concentrations of Ret siRNA or vehicle alone. (a) When podocytes were exposed to i-Fect alone, there was no toxicity. (a and b) A transfection efficiency of nearly 100% was achieved with 100 nM concentration of Ret siRNA (b). Co-transfection with a fluorescently tagged control siRNA was used to determine the transfection efficiency, and fluorescence microscopy revealed a perinuclear localization of th...
Source: siRNA and DsiRNA Transfection Efficiency - April 9, 2008 Category: Neuroscience Tags: Nearly 100% Transfection Efficiency Transfecting Podocytes Ret Source Type: organizations

RNAi therapeutics: a potential new class of pharmaceutical drugsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Comprehensive and Cogent overview on delivery methods:David Bumcrot1, Muthiah Manoharan1, Victor Koteliansky1 and Dinah W Y Sah1. RNAi therapeutics: a potential new class of pharmaceutical drugsNature Chemical Biology 2, 711-719 (2006) doi:10.1038/nchembio839
Source: siRNA and DsiRNA Transfection Efficiency - November 26, 2007 Category: Neuroscience Tags: Dinah Sah siRNAi Liposome siRNA Complexes Source Type: organizations

i-Fect and Posters @ SfNEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
This study is supported by NIH grant R01NS046785.Disclosures: M. Luo , None; D. Zhang, None; E. Zhang, None; Q. Chen, None; P. Ge, None; D. Sah, None; T. Vanderah, None; F. Porreca, None; J. Lai, None.Support:NIH grant R01NS046785[Authors]. [Abstract Title]. Program No. XXX.XX. 2007 Neuroscience Meeting Planner. San Diego, CA: Society for Neuroscience, 2007. Online.Program#/Poster#:509.6/PP9Title:Small interfering RNA-mediated selective knockdown of NTS2 receptors reverses neurotensin-induced analgesia in ratsLocation:San Diego Convention Center: Halls B-HPresentation Start/End Time:Monday, Nov 05, 2007, 2:00 PM - 3:00 PMA...
Source: siRNA and DsiRNA Transfection Efficiency - November 8, 2007 Category: Neuroscience Tags: siRNA Transfection i-Fect in vivo Source Type: organizations

More on Delivering siRNA in vivoEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We are excited about this upcoming presentation at Society for Neuroscience comference presesentation. Neuromics' i-Fect ™ reagent was used to deliver siRNA in vivo to silence the NTS2 gene.Program#/Poster#:509.6/PP9Title:Small interfering RNA-mediated selective knockdown of NTS2 receptors reverses neurotensin-induced analgesia in ratsLocation:San Diego Convention Center: Halls B-HPresentation Start/End Time:Monday, Nov 05, 2007, 2:00 PM - 3:00 PMAuthors:*L. DORE-SAVARD1,2, G. ROUSSY1, M.-A. DANSEREAU1, K. BELLEVILLE1, N. BEAUDET1, M. BEHLKE2, P. SARRET1;1Physiology and Biophysics, Univ. Sherbrooke, Sherbrooke, PQ, Canad...
Source: siRNA and DsiRNA Transfection Efficiency - October 30, 2007 Category: Neuroscience Tags: siRNA RNAi i-Fect iFect Source Type: organizations

i-Fect and Nav1.8 Gene Silencing in vivoEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Referenced in United States Patent 20070105806Link to patent: Free Patents Online[0241] The effect of siRNAs against Nav1.8, formulated with iFECT, on complete Freund's adjuvant-induced tactile hypersensitivity was evaluated in rats (FIG. 5). Adult male Sprague-Dawley rats received an injection of CFA (150 uL) into the hindpaw on day 0. siRNAs against Nav1.8 were then administered by intrathecal bolus to the lumbar region of the spinal cord on days 1, 2 and 3; specifically, for each bolus injection, 2 ug of siRNA was complexed with iFECT transfection reagent (Neuromics, Minneapolis Minn., USA) at a ratio of 1:4 (w:v) in a ...
Source: siRNA and DsiRNA Transfection Efficiency - May 23, 2007 Category: Neuroscience Source Type: organizations

Magnetically Enhanced siRNA TransfectionEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We have added a new set of products that magnetically drive transfection.MATra products are designed to enhance the transfection capabilties of our i-Fect and pn-Fect kits.The technique requires lease or purchase of the Universal Magnetic Plate AND for i-Fect and other siRNA transfection reagents, please also purchase the MATra-siRNA Reagent. For pn-Fect and other Nucleic Acid transfectants, you would buy the MATra-A Regeant.If your cells are in suspension, you will also need MATra-s-Immobilizer.Manual for All MATra ProductsMATra FAQsMagnet Assisted Transfection AnimationReferences:Publications:Bertram, J. (2006) MATra - M...
Source: siRNA and DsiRNA Transfection Efficiency - April 3, 2007 Category: Neuroscience Tags: siRNA RNAi Magnetics Transfection Source Type: organizations

Duration of siRNA studies-pain study exampleEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We are working with an investigator on using RNAi to relieve pain and his study prompted some discussion about the length of time before an effect is seen. It is important to note that these types of experiments take time to see an effect. In studying the pain response, the effect may take even longer. Dorn et al (Nucleic Acids Res. 2004 Mar 16;32) examined the ability of siRNA to relieve chronic neuropathic pain targeting P2X3 in vivo. They showed a significant effect on the ability of the siRNA to relieve the pain; however there was no significant effect until around 6 days post. Another study by Hemmings-Mieszczak e...
Source: siRNA and DsiRNA Transfection Efficiency - February 7, 2007 Category: Neuroscience Source Type: organizations

Transfecting Primary CellsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
We welcome any comments on you experiences with transfecting primary cell lines with cationic lipids and/or electroporation.Here're are interesting comments of transfecting primary cells from Dr. Mark Behlke, CSO of IDT:"Just wanted to give you a project update. We have good transfection conditions worked out for LTK cells and CHO cells now. Different reagents and different conditions proved to be optimal for each cell line.The LTK cells proved to be a real nightmare. We were testing out different siRNAs and had a weird problem that it seemed that none of the siRNA worked even a little, with the weird observation that our ...
Source: siRNA and DsiRNA Transfection Efficiency - January 15, 2007 Category: Neuroscience Source Type: organizations

Protocols for Delivering siRNA to NeuronsEmail this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
Here's a link to the protocol: Transfecting Schwann Cells with i-Fect and below are related data. Figure:si RNA-mediated suppression of target gene expression in Schwann cells. A, Detergent extracts of siNeg- or siGly1- transfected Schwann cells were digested with heparitinase and subjected to immunoblot analysis with anti-glypican-1 antibodies (top);aliquots of undigested extracts wereimmunoblotted with anti-actin antibodies (bottom) to verify equal sample loading. B, Cell surface expression of glypican-1 was assessed by immunofluorescent staining of transfected cells 48 h after transfection using anti-glypican- 1 antibod...
Source: siRNA and DsiRNA Transfection Efficiency - January 12, 2007 Category: Neuroscience Tags: i-Fect dsiRNA Source Type: organizations