What is SilenciX®
What is SilenciX® • New strategy for Knock-Down (KD) based on EBV vector • Main benefits: Efficiency - Validation performed by qPCR - KD level guarantee: >70% Specificity High Stability and long term culture: 3-12 months No off-target effects Non-viral genome integrity Safe Ready to Use (RTU) SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. How SilenciX® works The Epstein-Barr (EBV)-based vector is anchored to chromosomes and replicates as a replicon without any integration in the genome. Reinforced by optimal hygromycin selection, the number of vectors is perfectly maintained. SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. How SilenciX® works • • • • Replicative vector ensures genome integrity Capable of anchoring to chromosomes bringing stability Hygromycin resistance reinforces EBV replication H1-promotor expression gives efficient siRNA delivery SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. Why SilenciX® “Silence is a fence around wisdom” Proverb To have access to a powerful cellular model – fast! To study a physiological role of your protein in cellular model – fast! To screen and analyze new therapeutics in the early stage of their development – fast! To save time and money – fast! SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. Quantitative PCR (qPCR) Quality Controls Three independent cell lines silenced for each targeted gene of interest are established The knock-down efficiency is monitored with qPCR. The most efficient* knock-down (70-80%) is selected for your experiment The remaining cell lines can be sent upon request to analyze intermediate effects. *The efficiency depends on the gene of interest SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. SilenciX®: Efficiency 70-80% of knockdown leads to a very low remaining gene expression in SilenciX, compared to endogenous expression. SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. SilenciX: Comparison to siRNA and shRNA technologies siRNA Stability 2‐3days shRNA HeLa and/or all human and rodent cell lines Several month, Several month Start new experiment with different cell batch Efficiency Production time Price Reproducibility Robustness Integration and off‐target effect Additional reagents* Additional experiments SilenciX Start new experiment with the same cell batch: just thaw the vial >70% validation by qPCR Few days, controlling each experiment ~$1,200, control not included no 5 weeks $833 shRNA only Final cost ~$3,000 Yes, with experience no no no yes Required Protocol optimization Transfection, qPCR Required Protocol optimization Transfection, Selection, qPCR 2/3days $3,690 High reproducibility, Homogeneous population Highly robust no No additional reagents Ready to use (RTU) SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. SilenciX® : “silencing” DUBs Deubiquitylases (DUBs): major players in a variety of cellular processes such as cell cycle, DNA damage response (DDR), cell proliferation, cell growth and apoptosis Ready to supply: • USP7 HeLa SilenciX • USP9x HeLA SilenciX • USP14 HeLa SilenciX • USP19 HeLa SilenciX Don’t see your DUB? Custom cell lines are available. Custom production with all attached human and rodent cell lines compatible with lipofectamine transfection. SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. Target-specific SilenciX® cell lines Successfully silencing over 100 genes Example: DNA repair Base Excision Repair, Nucleotide Excision Repair, Mismatch Repair and Double-Stranded Breaks Breast Cancer Susceptibility and Fanconi Anemia SLX-00027 FancD2 HeLa SilenciX SLX-00028 BRCA2 HeLa SilenciX SLX-00041 BRCA1 HeLa SilenciX Non-Homologous End Joining SLX-00010 Ligase IV HeLa SilenciX SLX-00005 DNA-PKcs HeLa SilenciX SLX-00018 XRCC4 HeLa SilenciX SLX-00021 XLF HeLa SilenciX Xeroderma Pigmentosum SLX-00015 XPA HeLa SilenciX SLX-00016 XPC HeLa SilenciX SLX-00003 XPF HeLa SilenciX SLX-00133 XPG HeLa SilenciX Sensors/transducers SLX-00029 ATM HeLa SilenciX SLX-00025 ATR HeLa SilenciX SLX-00085 PARG HeLa SilenciX SLX-00008 KIN17 HeLa SilenciX Homologous Recombination SLX-00022 Rad54a HeLa SilenciX SLX-00124 Rad54b HeLa SilenciX Nucleotide Excision Repair SLX-00006 HR23A HeLa SilenciX SLX-00007 HR23B HeLa SilenciX SLX-00002 ERCC1 HeLa SilenciX SLX-00015 XPA HeLa SilenciX SLX-00016 XPC HeLa SilenciX MRN complex SLX-00013 Rad50 HeLa SilenciX SLX-00012 NBS12 HeLa SilenciX SLX-00011 MRE11 HeLa SilenciX Mismatch Repair SLX-00023 MSH2 HeLa SilenciX SLX-00052 PCNA HeLa SilenciX SLX-00089 FEN1 HeLa SilenciX Base Excision Repair SLX-00053 NEIL1 HeLa SilenciX SLX-00089 FEN1 HeLa SilenciX SLX-00019 PARP1 HeLa SilenciX SLX-00020 PARP2 HeLa SilenciX SLX-00018 XRCC1 HeLa SilenciX SLX-00009 Ligase III HeLa SilenciX SLX-00014 Ogg1 HeLa SilenciX Double-strand Breaks SLX-00005 DNA-PKcs HeLa SilenciX SLX-00021 XLF HeLa SilenciX SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. SilenciX® at work Examples of custom SilenciX cell lines SilenciX works in MRC5-V2 (human fibroblasts) and MCF7 (beast cancer cell line) with specific knockdown efficiency up to 80%. Other cell lines are compatible with SilenciX technology: HeLa: Human immortal cell line from cervical cancer cells HEK293: Embryonic Kidney 293 cell line Caco-2: Epithelial colorectal adenocarcinoma cells AC16: Proliferating cardiomyocyte cell line SH-SY5Y: Neuroblastoma cell line RKO: Colorectal carcinoma cell line HCT116: Colon Cancer cell Line U2OS: Osteosarcoma cell line B16F10: Murine melanoma cells CHO: Chinese hamster ovary cells XPA/XPC - zinc finger proteins involved in DNA excision repair KIN17- RNA-binding protein, involved in DNA replication and the cellular response to DNA damage MRE11 – DNA double-strand break repair protein SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA. SilenciX® : What do you get For each cell lines, you receive : One vial of target specific SilenciX® cells One vial of control SilenciX® cells (transfected with a nonrelevant shRNA sequence Specification sheets with qPCR quality controls and details of shRNA sequence Complete user manual SilenciX® is a registered trademark of tebu-bio, technology licensed from the CEA.
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