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pCpGfree-siRNA & pCpGfree-siRNADUO Description pCpGfree-siRNA and pCpGfree-siRNADUO are CpG-free plasmids that allow the expression of one or two shRNAs, respectively. The absence of CpG dinucleotides in their backbone makes these plasmids less immunogenic and insensitive to CpG methylation thus ideal for long lasting expression of shRNA(s) in vivo . pCpGfree-siRNA The pCpGfree-siRNA plasmid combines the backbone of the pCpGfree plasmid with the shRNA expression cassette of the psiRNA plasmid. pCpGfree-siRNA features the human 7SK promoter, in which all CpG dinucleotides have been removed. The activity of this RNA Pol III promoter is augmented by the addition of the enhancer of the murine CMV immediate-early promoter, a Pol II enhancer 1 . The enhancer has been modified to remove all CpGs. The pCpGfree-siRNA plasmid uses Bbs I, an unusual restriction enzyme that generates asymmetric cohesive overhangs that are not compatible with each other to eliminate the risk of self-ligation of the vector. A second cloning strategy can be chosen by using Acc 65I and Hind III restriction sites. The pCpGfree-siRNA plasmid exploits the white/blue selection system to facilitate the screening of recombinant clones. It features an EM7-LacZ a-peptide cassette between the cloning sites to allow the easy discrimination between blue parental and white recombinant clones. pCpGfree-siRNADUO The pCpGfree-siRNADUO plasmid contains two shRNA cassette driven by a CpG-free version of the human 7SK promoter. The murine CMV enhancer has been added to increase the activity of both 7SK promoters. The first cassette features Acc65 I / Hind III cloning sites and a LacZ apeptide bacterial expression cassette for white/blue selection in E. coli. The second cassette contains Bbs I / Bbs I cloning sites and a GUS bacterial expression cassette, another system that allows white/blue selection in E. coli. pCpGfree-siRNA and pCpGfree-siRNA-DUO are provided with LyoComp GT115, a lyophilized competent E. coli mutant strain permissive with the white and blue color selection from the LacZ and GUS systems. Contents and Storage The pCpGfree-siRNA and pCpGfree-siRNADUO plasmids are provided as 50 μg of lyophilized DNA in a kit that includes the following components: - 20 μg control plasmid - 1 vial of LyoComp GT115 - 10 μg of each sequencing primer - 4 pouches of Fast-Media ® Zeo X-Gal or, - 2 pouches of Fast-Media ® Zeo X-Gal and 2 pouches of Fast-Media ® Zeo X-Gluc Products are shipped at room temperature. Store Fast-Media ® pouches at room temperature. Store all other components at -20°C. Acc 65I Bbs I Important Licensing Information: These products are covered by a Limited Use License (See Terms and Conditions on page 164). By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Bbs I Hind III Bbs I PRODUCT QUANTITY CAT. CODE pCpGfree-siRNA 1 kit kcpgf-sirna pCpGfree-siRNADUO 1 kit kcpgf-sirna2 Related Products Acc 65I LyoComp GT115, page 43 Fast-Media ® Zeo-X-Gal, page 45 Zeocin , page 18 Fast-Media ® Zeo-X-Gluc, page 45 www.invivogen.com/cpgfree-plasmid 137 Bbs I Hind III CpG-FREE DNA 6
CpG-FREE DNA 6 CpG-Free Genes - Synthetic CpG-free Genes Many non-mammalian genes are widely used as reporter or suicide genes in molecular and cellular studies. However these genes are recognized as foreign DNA by the vertebrate host leading to a progressive decline of their expression. To circumvent this limitation, InvivoGen has synthesized new alleles of these genes completely devoid of CpG dinucleotides. These synthetic CpG-free genes display higher activity and lower immunogenicity than their wild-type counterparts. Description The DNA sequence of the CpG-free genes was modified by optimizing the codon usage, eliminating the CpG nucleotides and avoiding secondary DNA structures without changing the amino acid sequence of the wild type proteins. CpG-free genes were chemically synthesized by assembling large oligonucleotides. Sequence integrity was verified by double-stranded sequencing. CpG-free genes are provided in the pSELECT-zeo plasmid (see page 32). pSELECT-zeo is a mammalian expression plasmid selectable in E. coli and mammalian cells with Zeocin . Each CpG-free gene is flanked by a unique restriction site at the 5’ and 3’ end to facilitate its subcloning into another vector. Contents and Storage Each pSELECT-zeo- plasmid is provided as 20 μg of lyophilized DNA. Product is shipped at room temperature and should be stored at -20°C. Plasmid is stable up to one year when properly stored. Each plasmid is supplied with 4 pouches of E. coli Fast-Media ® Zeo (2 TB and 2 Agar, see pages 44-45). GENE DESCRIPTION Suicide Genes Fcy::fur* S. cerevisiae cytosine deaminase-uracil phosphoribosyl transferase fusion 33 20 μg psetz-fcyfur HSV1-tk Herpes simplex virus 1 (HSV1) thymidine kinase 121 20 μg psetz-hsv1tk HSV1-tk::Sh* HSV1 thymidine kinase-Zeocin resistance fusion 372 20 μg psetz-hsv1tksh Reporter Genes GFP::Sh* Green fluorescent protein-Zeocin resistance fusion 110 20 μg psetz-zgfpsh GFP::Bsr* NEW Green fluorescent protein-Blasticidin resistance fusion 110 20 μg psetz-zgfpbsr LacZ b-Galactosidase 289 20 μg psetz-lacz LacZnls b-Galactosidase with SV40 nuclear localization signal 289 20 μg psetz-lacznls LacZ::Sh* b-Galactosidase-Zeocin resistance fusion 341 20 μg psetz-laczsh Luc::Sh* Firefly luciferase-Zeocin resistance fusion 151 20 μg psetz-lucsh SEAP Mouse secreted embryonic alkaline phosphatase 65 20 μg psetz-mseap * Fusion gene 138 www.invivogen.com/cpgfree-genes Note: The pSELECT-zeo plasmid backbone contains CpG dinucleotides. CpGs IN NATIVE GENE QUANTITY CAT. CODE
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INNOVATION WITHIN REACH InvivoGen
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CONTENTS AT A GLANCE 1 CELL CULTURE
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4 CYTOKINE SIGNALING TABLE of CONTE
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Mycoplasma Detection & Elimination
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PlasmoTest Contents - HEK-Blue -2
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Primocin - The Antimicrobial Shiel
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SEAP Reporter Gene System Secreted
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HEK-Blue Detection - Real-Time Det
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Blasticidin S Description Blasticid
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LyoVec New formulation LyoVec is
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Poly-Arginine-HA Tagged Reprogrammi
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LENTI-Smart - Lentiviral Vector Pr
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Lentiviral Vector Production and Ce
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Antibody generation using pFUSE-CHI
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Engineering Fc regions for altered
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pBROAD & pWHERE - Optimized Vector
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pSELECT-Tag - Expression of a GFP-,
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pVIVO - Expression of Two Genes of
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pORF Description pORF plasmids feat
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Prom A-List - Human and Rodent Pro
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PROMOTER GENE TISSUE DISTRIBUTION S
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ChemiComp E. coli ChemiComp E. coli
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Fast-Media ® Preparation 1. Empty
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RIG-I-like Receptors 54 RLR & RLR-R
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Toll-Like Receptors Toll-Like Recep
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www.invivogen.com/innate-immunity 5
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IPAF and NAIP5 IPAF (NLRC4, CLAN/CA
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C-Type Lectin Receptors C-type lect
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pUNO pUNO-HA Toll-Like Receptors (T
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GENE NAME/ALIASES DESCRIPTION CAT.
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pDUO - Gene Associations Features a
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pZERO-TLR & pDeNy - Dominant Negati
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293/TLR & 293/NOD Clones 293/TLR Cl
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Response of HEK-Blue hTLR2 cells t
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Ramos-Blue Cells - NF-kB/AP-1 Repo
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MEF Cells C3H/TLR4mut & C3H/WT MEFs
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Antibodies for Detection ANTIBODY D
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- NF-kB binding site: Nuclear facto
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PRODUCT ORIGIN/DESCRIPTION TLR2 Ago
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PRODUCT ORIGIN/DESCRIPTION TLR9 Ago
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PRODUCT ORIGIN/DESCRIPTION Dectin-1
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LPS-PG (Porphyromonas gingivalis) R
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• FLA-ST ultrapure was purified b
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Page 90 and 91: 1. Hornung V. et al., 2006. 5'-Trip
Page 92 and 93: TLR/NOD Ligand Screening InvivoGen
Page 94 and 95: Signal Transduction Inhibitors Invi
Page 96 and 97: Pepinh-MYD - MyD88 inhibitory pepti
Page 98 and 99: 1. Martinon F,. et al., 2002. The i
Page 100 and 101: Nigericin - NLRP3 Inflammasome Indu
Page 102 and 103: Autophagy is a pathway by which cyt
Page 104 and 105: 4 CYTOKINE SIGNALING Cytokine Genes
Page 106 and 107: Blue Cytokine Reporter Cells Blue
Page 108 and 109: HEK-Blue IFN-a/b Cells - Human Typ
Page 110 and 111: HEK-Blue IL-6 Cells - IL6 Reporter
Page 112 and 113: HEK-Blue IL-1b Cells - IL-1b Repor
Page 114 and 115: HEK-Blue IL-33/IL-1b Cells - IL-33
Page 116 and 117: HEK-Blue CD40L Cells - CD40L Repor
Page 118 and 119: 5 ANTIBODIES & VACCINATION Antibodi
Page 120 and 121: Antibody generation using pFUSE-CHI
Page 122 and 123: Immunoglobulin A The mucosal surfac
Page 124 and 125: IgA Detection and Quantification In
Page 126 and 127: IgG Antibodies InvivoGen offers a s
Page 128 and 129: Mediators? 1. Leroux-Roels G., 2010
Page 130 and 131: Poly(I:C) Synthetic double-stranded
Page 132 and 133: pVAC - Induction of Neutralizing An
Page 134 and 135: CpG-Free DNA CpGs and the Immune Re
Page 136 and 137: pCpGfree-basic & pCpGfree-promoter
Page 140 and 141: Custom-Made CpG-Free Genes InvivoGe
Page 143 and 144: RNA INTERFERENCE 7 psiRNA System I
Page 145 and 146: RNA INTERFERENCE 7 psiRNA-h7SK - 7S
Page 147 and 148: RNA INTERFERENCE 7 psiTEST System
Page 149 and 150: RNA INTERFERENCE 7 Ready-Made psiRN
Page 151 and 152: RNA INTERFERENCE 7 Custom-Made psiR
Page 153 and 154: INHIBITORS 8 Inhibitors PRODUCT DES
Page 155 and 156: INHIBITORS 8 Pattern Recognition Re
Page 157 and 158: INHIBITORS 8 Triptolide - NF-kB Inh
Page 159 and 160: INHIBITORS 8 Heat Shock Protein Inh
Page 161 and 162: 9 CUSTOM SERVICES TLR/NOD Ligand Sc
Page 163 and 164: CUSTOM SERVICES 9 Custom Cloning :
Page 165 and 166: CUSTOM SERVICES 9 Prices 164 TERMS
Page 167 and 168: 166 PRODUCT (QUANTITY) CAT. CODE PA
Page 173 and 174: A 17-AAG 17-AAGH2 17-AEP-GA 2-Amino
Page 175 and 176: - ODN 4084-F - ODN INH-1 - ODN INH-
Page 177 and 178: ARGENTINA Genbiotech S.R.L. Tel: +5
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