MedWorm.com provides a medical RSS filtering service. Over 6000 RSS medical sources are combined and output via different filters. This feed contains the latest items from the 'siRNA and DsiRNA Transfection Efficiency' source. Tue, 07 Feb 2012 07:05:47 +0100 http://www.medworm.com/index.php?rid=5665100&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2012%2F02%2Fsirna-delivery-group-on-linkedin.html I wanted to make readers aware of an excellent discussion group on Linkedin named "siRNA Delivery". Included are tip, updates on commercialization and key publications. Here're some examples:Alnylam Announces Publication of Pre-clinical Results with ALN-HTT, an RNAi Therapeutic for the Treatment of Huntington’s Disease, in Experimental NeurologyDelivering siRNA to Neurons Life Technologies develops new drug delivery technology Recent experiments demonstrate a novel method of liposomal encapsulation of siRNA/RNA/DNA at 4C, could be a promising method to minimize degradation.Happy reading.    (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=5665100 Sat, 04 Feb 2012 16:44:00 +0100 5665100 http://www.medworm.com/index.php?rid=5513573&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F12%2Fdelivering-dna-to-cell-lines-and.html Neuromics is pleased to introduce DNA-FectTM and DNA-FectTM293 in vivo transfection reagents. These reagents deliver genes to various established cell lines as well as primary cells, which include HEK293, 293T, 293E, CHO, COS1, HeLa, NIH 3T3, insect cell lines (Sf9 and Sf21) and a variety of other eucaryotic cell lines with low cytotoxicity. GeneExpresso™ MAX reagent, 1.0 ml, is sufficient for 300 to 600 transfections in 24 well plates, or 150 to 300 transfections in 6 well plates.Capabilities include:• Proven to deliver DNA to difficult-to-transfect cells•Stable and easy to use•Suitable for high-thoroughput (HTS) applicationsImage: Comparing DNA-Fect vs Lipofectamine 2000 by FACS AnalysisDNA Delivery Protocol: (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=5513573 Thu, 15 Dec 2011 11:18:00 +0100 5513573 http://www.medworm.com/index.php?rid=5270952&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F09%2Farrestin-sirna-delivery-in-vivo-and.html Conclusions: We show here that intrathecal β-arrestin 2 siRNA in rats enhances analgesia and attenuates naloxone-induced withdrawal symptoms. This may warrant further investigation in the context of long-term use of intrathecal opioids for controlling chronic pain. (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=5270952 Thu, 29 Sep 2011 11:33:00 +0100 5270952 http://www.medworm.com/index.php?rid=5258259&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F09%2Fenhanced-transfection-efficiency-of.html In conclusion, modifications to the RT protocol of lipofection result in a significant and robust increase in the transfection efficiency of hES cells.Related Neuromics' Reagents for Testing and Gene Expression Analysis of Stem Cells:LumiSTEM™ in vitro Cell Based Assays-Designed for Primary Stem and Explanted Cells, Stem and Other Cell Lines.LUMENESC™ in vitro Cell Based Assays-Designed for Mesenchymal Stem/Stromal Cells (MSCs) and Cells derived from MSCs.HALO® in vitro Cell Based Assays-Designed for Lympho-Hematopoietic Stem and Progenitor Cells.Transfection Reagents (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=5258259 Tue, 27 Sep 2011 00:41:00 +0100 5258259 http://www.medworm.com/index.php?rid=5158147&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F08%2Fbidirectional-integrative-regulation-of.html I have reported use of our i-FectTM siRNA delivery kit for gene expression analysis studies of DOR, hTERT, The β3 subunit of the Na+,K+-ATPase, rSNSR1, NTS1. NAV1.8, , TRPV1, Survivin, Flaviviruses and more. I am pleased to add the Cav1.2 calcium channel to this growing list. Congratulations to Dr. Marc Landry for discovering the interplay between microRNA-miR-103 and this calcium channel: Alexandre Favereaux, Olivier Thoumine, Rabia Bouali-Benazzouz, Virginie Roques, Marie-Amélie Papon, Sherine Abdel Salam, Guillaume Drutel, Claire Léger, André Calas, Frédéric Nagy and Marc Landry. Bidirectional integrative regulation of Cav1.2 calcium channel by microRNA miR-103: role in pain. The EMBO Journal , (29 July 2011) | doi:10.1038/emboj.2011.249.Abstract: Chronic pain states are character... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=5158147 Sat, 20 Aug 2011 18:15:00 +0100 5158147 http://www.medworm.com/index.php?rid=5118259&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F08%2Fpei-transfection-and-implications-for.html This is a great article for undertanding the root causes of cytotoxicity caused by PEI-DNA Polyplexes.Giovanna Grandinetti, Nilesh P. Ingle, and Theresa M. Reineke. Interaction of Poly(ethylenimine)–DNA Polyplexes with Mitochondria: Implications for a Mechanism of Cytotoxicity. Mol. Pharmaceutics, Article ASAP. Publication Date (Web): June 23, 2011. Copyright © 2011 American Chemical Society.Poly(ethylenimine) (PEI) and PEI-based systems have been widely studied for use as nucleic acid delivery vehicles. However, many of these vehicles display high cytotoxicity, rendering them unfit for therapeutic use. By exploring the mechanisms that cause cytotoxicity, and through understanding structure–function relationships between polymers and intracellular interactions, nucleic acid delivery v... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=5118259 Wed, 10 Aug 2011 20:44:00 +0100 5118259 http://www.medworm.com/index.php?rid=4852630&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F05%2Fneuroscience-best-transfection.html I have multiple posting of customer success in transfecting siRNA and plasmids into neurons using Neuromics' i-FectTM and pn-FectTM...related publications.Here I post excellent data, images and testimonials provided by researchers using our Magnetic Assisted Transfection (MATraTM). Our goal is to continue our journey towards having the best practices for gene expression analysis studies in the CNS.With Magnet Assisted Transfection,  IBA/Neuromics offers a very gentle and potent tool for the transfection of many kinds of neuronal cells. Magnet Assisted Transfection is the ideal solution to overcome problems related to the study of complex and easily interrupted systems.Transfection of primary cortical neuronsExample 1 Embryonic cortical neurons were transfected with human NCAM.Emb... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=4852630 Sat, 21 May 2011 15:54:00 +0100 4852630 http://www.medworm.com/index.php?rid=4820501&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F05%2Fpkasirna-block-hyperalgesia.html I have reported use of our i-FectTM siRNA delivery kit for gene expression analysis studies of DOR, hTERT, The β3 subunit of the Na+,K+-ATPase, rSNSR1, NTS1. NAV1.8, , TRPV1, Survivin, Flaviviruses and more. The data in this pub indicates that selective knock-down of spinal PKA activity by intrathecal (i.th.) pretreatment of rats with a PKA-selective small interference RNA (siRNA) mixture significantly attenuates sustained morphine-mediated augmentation of spinal CGRP immunoreactivity, thermal hyperalgesia, mechanical allodynia and antinociceptive tolerance. The present findings indicate that sustained morphine-mediated activation of spinal cAMP/PKA-dependent signaling may play an important role in opioid induced hyperalgesia: S. Tumati, W.R. Roeskea, T.M. Largent-Milnesa, T.W. Vanderaha,... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=4820501 Fri, 13 May 2011 18:00:00 +0100 4820501 http://www.medworm.com/index.php?rid=4747406&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F04%2Fasics-and-surgical-pain.html Dr. Eric Lingueglia, an INSERM group leader, and his team at the CNRS IPMC/IN2M have been doing impressive research using our i-Fect ™ siRNA Transfection Kits to study the role of Acid-Sensing Ion Channels in Postoperative Pain. The etiology and pathophysiology of this pain is poorly understood. Their work is shedding light on potential root causes: Emmanuel Deval, Jacques Noël, Xavier Gasull1, Anne Delaunay, Abdelkrim Alloui, Valérie Friend, Alain Eschalier, Michel Lazdunski, and Eric Lingueglia. Acid-Sensing Ion Channels in Postoperative Pain. The Journal of Neuroscience, 20 April 2011, 31(16): 6059-6066; doi: 10.1523/​JNEUROSCI.5266-10.2011....Ten microliters of a siRNA (2 μg)/i-Fect (Neuromics) mix was injected intrathecally between the L4 and L5 vertebrae of rats using a H... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=4747406 Mon, 25 Apr 2011 14:01:00 +0100 4747406 http://www.medworm.com/index.php?rid=4723451&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2011%2F04%2Fdelivering-trpv1-shrna-to-drg-of-t8-l3.html Conclusion: Our data show that selective knockdown of TRPV1 expressed in DRG of T8-L3 segments of the spinal cord and their central and peripheral terminals increases blood pressure, suggesting that neuronal TRPV1 in these segments possesses a tonic anti-hypertensive effect possibly via suppression of the sympathetic nerve activity. (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=4723451 Sun, 17 Apr 2011 17:50:00 +0100 4723451 http://www.medworm.com/index.php?rid=3980550&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F09%2Ftransfecting-primary-cortical-neurons.html Harnessing the power of MATraTM (Magnetic Assisted) Transfection Kits.Background: The neural cell adhesion molecule (NCAM) plays a major role during development of the nervous system and in synapse plasticity in the adult brain (Diestel et al., 2007). Many studies provide evidence that NCAM can regulate processes like cell migration, axon growth and fasciculation. Endocytosis of NCAM might play a decisive role in these processes as it can potentially enable a quick change in cell adhesion between the cells or towards the extracellular matrix. Endocytosis ofNCAM might also influence these processes by activating specific signal transduction pathways.Primary cortical neurons present a good in vitro system for these investigations since they allow analysis of molecules within growth cones. Fo... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3980550 Sat, 18 Sep 2010 04:01:00 +0100 3980550 http://www.medworm.com/index.php?rid=3902742&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F08%2Fi-fect-and-sirna-delvery-to-toll-like.html Conclusions/Significance: These findings suggest that suppression of TLR4 mediated by intrathecally administered siRNA may be a new strategy for the treatment of neuropathic pain.Images: Screening siRNA for an efficient suppression of TLR4 expression in vitro. HEK-293 cells were co-transfected with both pEGFRC1-TLR4 and either one of three independent siRNA oligonucleotides targeting TLR4 (TLR4-siRNA1-3) or a control siRNA (MM-siRNA). Two days after transfection, EGFP fluorescence was observed under microscope (A) or quantified by flow cytometry (B). (A) EGFP fluorescence under an inverted fluorescence microscope (×100) or cell density under an optical microscope (×100). A, control; B, siRNA1; C, siRNA2; D, siRNA3. (B) The quantification of TLR4-EGFP fluorescence intensity upon siRNA kno... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3902742 Wed, 25 Aug 2010 11:32:00 +0100 3902742 http://www.medworm.com/index.php?rid=3872330&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F08%2Fintra-i-fect-and-intravenous-delivery.html Deliver siRNA in-vivo with stunning results! Introductory Special-200 to 600 USD (valid through 9/30/2010)These intra-i-Fect kits are designed to deliver siRNA in vivo via intravenous injections with high efficiency to specific tissue in rats and mice. The protocol involves these simple steps: prep, mix, dry, hydrate and inject.Figure: siRNAs knock down profiles of the gene related to cancer, diabetes, obesity, steatosis hepatitis, cirrhosis and a gene specifically expressed in endothelial cells in liverThey are developed using a proprietary platform that uses nano-particles as the delivery vehicle. This platform enables:•Effective delivery (60%+ knockdown) with no toxicity.•Scalable to high throughput siRNA based gene screening.•Consistent and reproducible results. Protocolsintra-i-... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3872330 Mon, 16 Aug 2010 17:57:00 +0100 3872330 http://www.medworm.com/index.php?rid=3848728&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F08%2Fi-fect-survivin-and-gliobastomas.html I would like to add Survivin to the list of genes successfully silenced in-vitro and in-vivo using our i-FectTMsiRNA delivery kit.The list includes: DOR, hTERT, The β3 subunit of the Na+,K+-ATPase, rSNSR1, NTS1. NAV1.8 and moreJoseph George, Naren L. Banik and Swapan K. Ray. Survivin knockdown and concurrent 4-HPR treatment controlled human glioblastoma in vitro and in vivo. Neuro-Oncology, doi:10.1093/neuonc/noq079....survivin siRNA cDNA was suspended in RNAse free sterile water (25 μg DNA/10 μl) and mixed (1:4 v/v) with i-Fect transfection reagent (Neuromics)...Delivery of the Surivivin siRNA resulted in significant decreases in Glioblatoma Tumor Size. (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3848728 Mon, 09 Aug 2010 18:19:00 +0100 3848728 http://www.medworm.com/index.php?rid=3710397&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F06%2Fusing-matra-for-sirna-transfection-of.html MATRaTM -Magnet Assisted Transfection is an easy-to-handle, very fast and highly efficient technology to transfect cells in culture with siRNA. Multiple successes with the system includes Carcinoma Cell Lines. Efficient transient transfection of siRNA in head and neck cancer cells. The cell line ANT-1 was transiently transfected with MATra-A (1 µl/1 µg DNA) in a 6 well format (5 x 105 cells/cavity) with siRNA against protein 1 (100 nM). After 24 hours total RNA was isolated and expression of protein 1-specific mRNA determined by RT-PCR (upper lane). SiRNA 13 are three different oligonucleotide sequences. Control for consistent loading and cDNA quality: expression of ubiquitary GAPDH mRNA (lower lane).Protein 2 expression in head and neck cancer cells GHD-1. GHD-1 cells (5 x 105 cells/cav... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3710397 Tue, 29 Jun 2010 12:21:00 +0100 3710397 http://www.medworm.com/index.php?rid=3699326&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F06%2Fsirna-and-i-fect-for-study-of-retinal.html In this study, researchers used i-Fect to transfect and immortalized cell line from Mouse cones (661W) expressing ELOVL4. Using siRNA designed to silence the ELOVL4 gene, they used i-Fect + siRNA to transfect cells cultured at a density of 2X105. Knockdown was achieved as confirmed by western blot analysis. (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3699326 Fri, 25 Jun 2010 23:42:00 +0100 3699326 http://www.medworm.com/index.php?rid=3592109&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F05%2Fraf-1-selective-sirna-and-response-to.html Conclusions and implications: Raf-1 played a significant role in sustained morphine-mediated paradoxical pain sensitization and antinociceptive tolerance in vivo. These findings suggest novel pharmacological approaches to improve the long-term utility of opioids in the treatment of chronic pain. (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3592109 Mon, 24 May 2010 01:03:00 +0100 3592109 http://www.medworm.com/index.php?rid=3545358&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F05%2Fandy-miller-pesentation-wow.html Deliverying siRNA in vivo is faced with may challenges. Non specific and immune responses are at the top of the list. Researchers are fighting innate biology when successfully delivery.That said, I just came across an excellent presentation. Creative chemistry indeed! Here's the link: http://www.labtube.tv/avc-interest.aspx?i=5&c=1&v=327.Andy Miller talking at RNAi Europe 2009Andy Miller from Imperial College London giving his keynote lecture at RNAi Europe in Berlin. The talk was entitled, 'Enabling RNAi Therapeutics with Safe, Synthetic, Self Assembling Nanoparticles'.Published : 2009/10/27 (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3545358 Sat, 08 May 2010 02:46:00 +0100 3545358 http://www.medworm.com/index.php?rid=3529553&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F05%2Frnainet-check-it-out.html RNAi.net is a portal that has done an excellent job at providing a gateway to many resources to help researchers using siRNAs for gene expression analysis.There webcast link is particulary useful. Included is a presentation by one of our collaborators: Dr. Mark Behlke. Here's the abstract:Dicer-substrate siRNAs (DsiRNAs) are synthetic oligonucleotides that are processed by Dicer prior to RISC loading. DsiRNAs often show improved potency over traditional siRNAs in vitro and can have similar benefits in vivo. In collaboration with Dicerna Pharmaceuticals, systematic high throughput screening of DsiRNAs is in progress to identify ultra-potent sites in pharmaceutically relevant target genes. The results of a KRAS screening project will be discussed where over 400 synthetic siRNAs were tested i... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3529553 Tue, 04 May 2010 09:35:00 +0100 3529553 http://www.medworm.com/index.php?rid=3432651&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F04%2Fdirect-application-of-sirna-for-in-vivo.html My friends at McGill University have recently published in depth methods for using siRNA to study pain. Dr. Philippe Sarret have done extensive work delivering siRNA + i-FectTM in vivo for gene expression analysis of specific pain receptors.Here's a link to the book chapter from Springer Protocols:25. Direct Application of siRNA for In Vivo Pain ResearchBy: Philippe Sarret , Louis Doré-Savard, Nicolas BeaudetAffiliation(s): (1) Department of Physiology and Biophysics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, QC, CanadaBook Title: RNA Interference: From Biology to Clinical ApplicationsSeries: Methods in Molecular Biology Volume: 623 Pub. Date: May-01-2010 Page Range: 383-395 DOI: 10.1007/978-1-60761-588-0_25Abstract: Pain is the new burden of the twen... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3432651 Thu, 01 Apr 2010 21:48:00 +0100 3432651 http://www.medworm.com/index.php?rid=3287514&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F02%2Fi-fect-and-more-knockdown-success.html In this study researchers showed specific knockdown of CaV1.2 in the spinal dorsal horn reversed the neuropathy-associated mechanical hypersensitivity and the hyperexcitability and increased responsiveness of dorsal horn neurons. Intrathecal application of anti-CaV1.2 siRNAs confirmed the preceding results.Here's a link to the related pub: Pascal Fossat, Eric Dobremez, Rabia Bouali-Benazzouz, Alexandre Favereaux, Sandrine S. Bertrand, Kalle Kilk, Claire Léger, Jean-René Cazalets, Ülo Langel, Marc Landry and Frédéric Nagy. Knockdown of L Calcium Channel Subtypes: Differential Effects in Neuropathic Pain. The Journal of Neuroscience, January 20, 2010, 30(3):1073-1085; doi:10.1523/JNEUROSCI.3145-09.2010We used siRNA targeting several splice variants of CaV1.2 ("Silencer Select Pre-design... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3287514 Thu, 18 Feb 2010 18:14:00 +0100 3287514 http://www.medworm.com/index.php?rid=3153163&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2010%2F01%2Fdelivering-sirna-in-mice-for-studying.html Researchers have successfully delivered siRNA in-vitro and in-vivo using Neuromics' i-Fect ™ siRNA Transfection Reagent. Gene expression studies include: DOR, hTERT, The β3 subunit of the Na+,K+-ATPase, rSNSR1, NTS1. NAV1.8 and more.Here's a link to all transfection publications: Transfection Kit PubsWe are pleased to present yet another study and related publication. This includes one of the first successful delivery of siRNA in mice using i-Fect ™ :Yan Chen, Cheng Yang, and Zaijie Jim Wang. Ca2+/Calmodulin-Dependent Protein Kinase II Is Required for the Initiation and Maintenance of Opioid-Induced Hyperalgesia. The Journal of Neuroscience, January 6, 2010, 30(1):38-46; doi:10.1523/JNEUROSCI.4346-09.2010....KN93 and KN92 were administered intrathecally by percutaneous puncture throug... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3153163 Thu, 07 Jan 2010 17:18:00 +0100 3153163 http://www.medworm.com/index.php?rid=3047938&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2009%2F12%2Fusing-i-fect-for-treatment-of.html Conclusions: Our study showed that the combination of hTERT siRNA and IFN-γ effectively inhibited angiogenesis and tumor progression through the downregulation of molecules involved in these processes. Therefore, the combination of hTERT siRNA and IFN-γ is a promising therapeutic strategy for controlling the growth of human glioblastoma. (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=3047938 Tue, 01 Dec 2009 22:14:00 +0100 3047938 http://www.medworm.com/index.php?rid=2508326&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2009%2F06%2Fdelivering-27mer-dsirnas-to-mice-drgs.html 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); Jul;16(7):1331-9. Epub 2008 Jun 3 doi:10.1038/mt.2008.98.Using ultra low dose of DsiRNAs complexed with Neuromics’ i-Fect , they were able to successfully reduce NTS2... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=2508326 Tue, 23 Jun 2009 15:39:00 +0100 2508326 http://www.medworm.com/index.php?rid=2347272&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2009%2F04%2Fknockdown-of-rsnsr1-in-vivo.html 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 administration by mixing the RNA solution (200 uM in annealing buffer) with the transfection reagent i-FectTM (Neuromics) at a ratio of 1:4 (w:v) for a final siRNA/ lipid complex c... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=2347272 Mon, 20 Apr 2009 15:12:00 +0100 2347272 http://www.medworm.com/index.php?rid=2330128&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2009%2F04%2Fdid-your-rnai-experiment-work.html 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) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=2330128 Mon, 06 Apr 2009 14:37:00 +0100 2330128 http://www.medworm.com/index.php?rid=2042597&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F12%2Fintrathecal-delivery-of-sirna.html 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). After recovery from the surgery (5–7 days), the animals received intrathecal injections (2 ug siRNA/10 ul/rat) of either a lipid encapsulated Raf-1-selective siRNA mix... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=2042597 Tue, 16 Dec 2008 19:44:00 +0100 2042597 http://www.medworm.com/index.php?rid=1926872&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F11%2Fdelivering-naked-sirna-by-direct.html 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) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1926872 Sun, 02 Nov 2008 15:00:00 +0100 1926872 http://www.medworm.com/index.php?rid=1889287&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F10%2Facic3-receptors-knockdown-in-vivo.html 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 by MWG Biotech (Germany) were injected in rats by intrathecal bolus to the lumbar region of the spinal cord once a day for 3 days before the induction of inflammation wi... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1889287 Sun, 19 Oct 2008 19:10:00 +0100 1889287 http://www.medworm.com/index.php?rid=1851825&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F10%2Fsirna-mediated-gene-silencing.html 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 (using mini-osmotic pumps)Validation We will continue to track advances by Dr. Lai and team. (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1851825 Sat, 04 Oct 2008 14:08:00 +0100 1851825 http://www.medworm.com/index.php?rid=1740469&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F08%2Ftransfecting-primary-neurons.html 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 have published results from there studies of C2AP and Cbl-3/Cbl-c and Ret Transduction. One of the keys to this study was using siRNA to silence CD2AP and Cbl-3 expression. ... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1740469 Thu, 28 Aug 2008 21:50:00 +0100 1740469 http://www.medworm.com/index.php?rid=1730898&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F08%2Fdown-regulating-smad-and-neuro.html 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. Vehicle only control rats undergo the same treatment but are injected with an equal volume of 0.09% saline in a tail vein. At the same treatment time points as the SB-505124... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1730898 Mon, 25 Aug 2008 05:51:00 +0100 1730898 http://www.medworm.com/index.php?rid=1672506&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F07%2Fsirna-and-diabetes.html 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) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1672506 Fri, 01 Aug 2008 01:19:00 +0100 1672506 http://www.medworm.com/index.php?rid=1650297&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F07%2Fimproving-27mer-dsirna-preformance.html 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 is an important publication for researchers wanting to better understand: The mechanisms behind successful delivery of DsiRNA for gene expression studies.Variations in po... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650297 Mon, 21 Jul 2008 02:53:00 +0100 1650297 http://www.medworm.com/index.php?rid=1650298&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F07%2Fcoming-soon.html 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) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650298 Tue, 08 Jul 2008 14:20:00 +0100 1650298 http://www.medworm.com/index.php?rid=1650299&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F06%2Frnai-researchers-galvanized-by-advances.html 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 interferon activation, however we now know that short RNA duplexes can be safely used in mammalian systems both in vitro and in vivo. The technology has rapidly matured, thanks i... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650299 Wed, 18 Jun 2008 14:33:00 +0100 1650299 http://www.medworm.com/index.php?rid=1650300&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F06%2Fhi-thru-put-delivery-of-sirnadsirna.html NEW PRODUCTi-Fect ™ Hi Put 96-NewLearn about transfection products @ neuromics (Source: siRNA and DsiRNA Transfection Efficiency) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650300 Mon, 16 Jun 2008 14:58:00 +0100 1650300 http://www.medworm.com/index.php?rid=1650301&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F06%2Fcentral-delivery-of-dsirna.html 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). As seen by confocal microscopy, the staining is not uniformly distributed among the cells. Higher-magnification images also show that the fluorescent signal is detecte... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650301 Wed, 04 Jun 2008 19:53:00 +0100 1650301 http://www.medworm.com/index.php?rid=1650302&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2008%2F04%2Fret-knockdown-using-small-interference.html 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 the tagged RNA (b, arrowhead). (B) Western blot analysis of Ret after transfection: Ret immunoblotting (top) of WCL 2, 3, or 4 d after transfection revealed that Ret was do... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650302 Wed, 09 Apr 2008 15:30:00 +0100 1650302 http://www.medworm.com/index.php?rid=1650303&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F11%2Frnai-therapeutics-potential-new-class.html 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) siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650303 Mon, 26 Nov 2007 17:27:00 +0100 1650303 http://www.medworm.com/index.php?rid=1650304&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F11%2Fi-fect-and-posters-sfn.html 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 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... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650304 Thu, 08 Nov 2007 01:44:00 +0100 1650304 http://www.medworm.com/index.php?rid=1650305&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F10%2Fmore-on-delivering-sirna-in-vivo.html 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, Canada; 2Integrated DNA Technologies Inc., Coralville, IA2007Copyright by the Society for Neuroscience all rights reserved. Permission to republish any abstract or part of any... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650305 Tue, 30 Oct 2007 12:29:00 +0100 1650305 http://www.medworm.com/index.php?rid=1650306&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F05%2Fi-fect-and-silencing-nav18-gene.html 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 total volume of 10 uL. Five groups of rats (with 5 rats per group) were treated with either siRNA (AL-DP-6049, AL-DP-6209, AL-DP-6217 or AL-DP-6218; Table 1), or PBS, in ... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650306 Wed, 23 May 2007 00:07:00 +0100 1650306 http://www.medworm.com/index.php?rid=1650307&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F04%2Fincreasing-sirna-transfection.html 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 - Magnet Assisted Transfection: Combining Nanotechnology and Magnetic Forces to Improve Intracellular Delivery of Nucleic Acids. Current Pharmaceutical Biotechnology 7, 277-... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650307 Mon, 02 Apr 2007 23:44:00 +0100 1650307 http://www.medworm.com/index.php?rid=1650308&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F02%2Fduration-of-sirna-studies-pain-study.html 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 et al (Nucleic Acids Research, 2003, Vol. 31, No. 8 2117-2126) showed that in an in vitro model, oligofectamine-mediated transfection of siRNA resulted in 60±90% downregu... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650308 Wed, 07 Feb 2007 04:07:00 +0100 1650308 http://www.medworm.com/index.php?rid=1650309&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F01%2Ftransfecting-primary-cells.html 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 controls were “very low”. After quite a bit of investigation, we finally figured out that the LTK cells were triggering an IFN pathway response to the siRNAs. Unfortu... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650309 Mon, 15 Jan 2007 19:18:00 +0100 1650309 http://www.medworm.com/index.php?rid=1650310&cid=s_37786_168_f&fid=37786&url=http%3A%2F%2Fsirnatransfection.blogspot.com%2F2007%2F01%2Fprotocols-for-delivering-sirna-to.html 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 antibodies (green) and DAPI (4',6'-diamidino-2 phenylindole) to stain nuclei (red). C, Schwann cells were transfected with siNeg or si 4(V)and conditioned media and cell lysates... siRNA and DsiRNA Transfection Efficiency news http://www.medworm.com/rss/comments.php?id=1650310 Fri, 12 Jan 2007 22:33:00 +0100 1650310