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Transfection and Genome Engineering Innovation Delivered Transfection Selection Guide 4 Contents Transfection Decision Tree 5 Superior Transfection Reagents 6

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Transfection and Genome Engineering Innovation Delivered Transfection Selection Guide 4 Contents Transfection Decision Tree 5 Superior Transfection Reagents 6 Transfection and genome engineering Innovation delivered Transfection selection guide 4 Contents Transfection decision tree 5 Superior transfection reagents 6 Lipofectamine® 3000 Transfection Reagent 6 Lipofectamine® RNAiMAX Transfection Reagent 10 Lipofectamine® MessengerMAX™ Transfection Reagent 11 ExpiFectamine™ 293 Transfection Kit 14 Broad-spectrum transfection reagent 15 Lipofectamine® 2000 Transfection Reagent 15 Electroporation 16 Neon® Transfection System 16 In vivo delivery 17 Invivofectamine 3.0 Reagent 17 Transfection-related products 19 Opti-MEM® I Reduced-Serum Medium 19 Selection antibiotics 19 Genome modulation and engineering 20 Ambion® siRNA 20 mirVana™ miRNA Mimics and Inhibitors 22 GeneArt® CRISPR tools 24 GeneArt® TAL effector tools 25 Ordering information 26 50,000 Transfection decision tree Transfection selection guide 45,000 Transfection Transfection 40,000 decision tree 35,000 Transfection is the process by which nucleic acids are introduced into eukaryotic cells. 30,000 PAYLOAD RNAi Plasmid DNA mRNA Techniques vary widely and include lipid-based transfection and physical methods such as 25,000 ® siRNA, miRNA Cas9 DNA, GFP, Luc, shRNA Cas9 mRNA, GFP mRNA electroporation. Lipofectamine transfection reagents are the most trusted and cited in the 20,000 scientific literature due to their superior transfection performance and broad cell spectrum. 15,000 An overview of our most effective transfection products is shown in Table 1 to help you 10,000 COTRANSFECTION choose the solution that’s right for you. 5,000 0 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 201020112012 2013 2014 Table 1. Transfection selection guide—more blocks represent higher Cumulative number of publications citing the use of Lipofectamine® family of reagents since 1996. ® ® ® Lipofectamine® transfection efficiency into a greater number of cell types. Lipofectamine Lipofectamine Lipofectamine TRANSFECTION TM Difficult-to- REAGENT RNAiMAX 2000, 3000 3000 MessengerMAX Co- Easy-to-transfect transfect Primary Suspension Transfection product DNA mRNA RNAi delivery* cells cells cells Stem cells cells Superior transfection reagents Lipofectamine® 3000 Transfection Reagent Lipofectamine® RNAiMAX Transfection Reagent GREAT COULD BE POOR GREAT COULD BE POOR Lipofectamine® MessengerMAX™ RESULTS BETTER RESULTS RESULTS BETTER RESULTS Transfection Reagent ExpiFectamine™ 293 ™ Transfection Kit Recommended for Expi293F cells Broad-spectrum transfection reagent Lipofectamine® 2000 Transfection Reagent TRY TRY TRY TRY ® ® Electroporation 1 Reverse transfection 1 Neon Transfection 1 Different DNA 1 mRNA + Lipofectamine 2 Different cell density System concentration MessengerMAX™ reagent Neon® Transfection System 3 Increased siRNA 2 Viruses with shRNA 2 Different cell density 2 Plasmid DNA + Neon® concentration Transfection System In vivo delivery 3 mRNA + Neon® Transfection System Invivofectamine 3.0 Reagent In vivo delivery to liver following tail vein injection ® Neon 4 Viruses Transfection System *Cotransfection of RNAi vector and siRNA. 4 5 Gentle with low toxicity Enhance your cancer research ® Superior transfection reagents Lipofectamine 3000 reagent was designed to optimize every step The superior transfection efficiency and broad cell spectrum of Superior in the transfection process. The superior transfection performance Lipofectamine® 3000 reagent enables excellent flexibility to use your allows you to reduce the reagent dose and improve cell viability cell line of choice in cancer studies (Figure 4). transfection reagents ® Lipofectamine 3000 Transfection Reagent “We were very happy and surprised to see when working with your cell line of interest (Figure 3). ® Lipofectamine® 3000 reagent leverages our most advanced lipid Lipofectamine 3000 (reagent) provide a nanoparticle technology to enable superior transfection performance more than 10-fold difference in transfection and reproducible results. It delivers exceptional transfection efficiency efficiency in our difficult-to-transfect cell line. into the widest range of difficult-to-transfect and common cell types There was even a reduction in cell death. (Figures 1 and 2) with improved cell viability. Awesome results!” – Rui Eduardo Castro, PhD % Transfection University of Lisbon 100 Transfection efciency in cancer cell line panel Lipofectamine® 3000 reagent offers: 90 100 • Superior transfection efficiency—into the broadest spectrum of Lipofectamine® 3000 Lipofectamine® 3000 Lipofectamine® 2000 Competitor y 80 c difficult-to-transfect cell types ® n 80 Lipofectamine 2000 e i 70 c • Improved cell viability—gentle on your cells, with low toxicity f e 60 HEK 293 60 on i t 40 ec 50 f s n a 40 r 20 T HeLa % Transfection 30 % Lipofectamine® 2000 0 5 7 1 0 0 9 0 3 6 8 T 1 3 2 1 8 D - - - - T L 2 i ® 2 R 20 s - 14 V 48 62 4 54 46 Lipofectamine 3000 46 H - - o - L 193 ak - 578 H O I W a W T - E B - - C I s ® ANC S S S B DU K M Lipofectamine LTX M P H 10 NC - - S HCC LNCaP NC K S DA 0 M 0.1 0.2 0.3 0.4 Cell line Dose (µL) Western blot, HepG2 HepG2 Lipofectamine® Lipofectamine® Competitor Competitor 2000 3000 1 2 Figure 3. Superior transfection efficiency of Lipofectamine® 3000 Figure 4. Transfection efficiency in a cancer cell line panel. Lipofectamine® reagent at low doses. Each reagent was used to transfect HeLa cells in a 2000 reagent and Lipofectamine® 3000 reagent were used to transfect 17 cell GST-STAT 96-well format at the indicated doses with an emerald green fluorescent lines with a GFP-expressing plasmid in a 24-well plate format using 0.5 µg A549 protein (emGFP)–expressing vector. Analysis was performed 48 hours plasmid/well and the recommended protocols for each reagent. GFP expression posttransfection using flow cytometry to determine percent transfection was analyzed 48 hours posttransfection. Each condition was tested in triplicate, ® β-actin efficiency. Lipofectamine 3000 reagent delivered higher efficiency than both and the data points with error bars show mean transfection efficiency with Lipofectamine® 2000 reagent and Lipofectamine® LTX reagent. standard deviation. Figure 2. Lipofectamine® 3000 reagent outperforms Lipofectamine® 2000 and competitor reagents. Each reagent was used to transfect HEK 293, Figure 1. Enhanced protein expression using Lipofectamine® 3000 reagent. HeLa, LNCaP, HepG2, and A549 cell lines in a 96-well format, and green HepG2 cells were transfected with a vector expressing GST-tagged STAT fluorescent protein (GFP) expression was analyzed 48 hours posttransfection. protein. A western blot was performed to determine the level of expression using Lipofectamine® 3000 reagent provided higher GFP transfection efficiency than β-actin as a control. Lipofectamine® 2000 and competitor reagents for all five cell lines. 6 7 Generate induced pluripotent stem cells Improve gene editing outcomes Transfect stem cells ® ® Induced pluripotent stem cells (iPSCs) hold immense promise for Lipofectamine 3000 reagent was developed to break through Lipofectamine 3000 reagent was developed to unleash the power Superior the future of regenerative medicine and personalized therapeutic the boundaries of traditional delivery methods and facilitate new of stem cells by providing a highly efficient, cost-effective nucleic treatments for a myriad of diseases and conditions. The superior technologies, such as genome engineering, in more biologically acid delivery alternative to electroporation (Figure 7). This advanced transfection reagents transfection efficiency of Lipofectamine® 3000 reagent in relevant systems. With this reagent, GeneArt® CRISPR vectors lipid nanoparticle technology minimizes the stress on cells caused conjunction with the Epi5™ Episomal iPSC Reprogramming Kit targeting the AAVS1 locus in HepG2 and U2OS cells show improved by electroporation, simplifies the reprogramming workflow, and enables highly efficient reprogramming of somatic cells without the transfection efficiency (Figure 6), mean fluorescence intensity, enables advanced gene-editing technologies. need for electroporation (Figure 5). and genomic cleavage. High transfection and genome editing efficiency is also observed with GeneArt® Precision TALs. These advancements in delivery help minimize painstaking downstream workflows, enable easier stem cell manipulation, and enhance site- specific insertion of transgenes into the genome. A B A B 150,000 A B 200,000 150,000 100,000 intensity intensity P P 100,000 50,000 50,000 Mean OF Mean OF 0 0 DNA: 1.0 µg DNA: 1.3 µg Lipofectamine® 3000: 1.5 µL Lipofectamine® 3000: 1.5 µL SSEA4+/GFP+: 42% SSEA4+/GFP+: 69% 3 µL Lipofectamine ® 2000 1.5 µL Lipofectamine ® 3000 3 µL Lipofectamine ® 2000 1.5 µL Lipofectamine ® 3000 GFP MFI 247344 GFP MFI 456741 Figure 5. Reprogramming efficiency of Lipofectamine® 3000 reagent Figure 6. Transfection efficiency and protein expression using GeneArt® Figure 7. Transfection of stem cells. (A) H9 embryonic stem cells (ESCs) or (B) compared to electroporation. BJ fibroblasts as well as neonatal (HDFn) CRISPR Nuclease Vector. The vector contained an OFP reporter gene and was iPSCs were transfected using Lipofectamine® 3000 reagent. Cells were visualized and adult (HDFa) human dermal fibroblasts were reprogrammed to iPSCs by transfected with Lipofectamine® 2000 or Lipofectamine® 3000 reagent into (A) by fluorescence microscopy and processed using flow cytometry to determine transfection of Epi5™ vectors using either Lipofectamine® 3000 reagent or U2OS and (B) HepG2 cell lines. Bar graphs show reporter gene expression;
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