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optimized crispr design tool  (Synthego Inc)

 
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    Synthego Inc optimized crispr design tool
    Genetic ablation of ERAD sensitizes HepG2 cells to TNFα– and IL-12–induced cell death. A, diagram <t>illustrating</t> <t>CRISPR/Cas9-mediated</t> editing of the Sel1L gene in HepG2 cells. F, R1, and R2 represent PCR primers used for genotyping of Sel1L-edited cells. Black bars represent exons. B , top , PCR analysis of Sel1L-normal (Sel1L +/+ ) and deleted (Sel1L −/− ) cells. Bottom, Western blotting analysis of SEL1L expression in Sel1L +/+ and Sel1L −/− cells; the upper band is a nonspecific band. C, average cell death level of Sel1L +/+ and Sel1L −/− cells as indicated by the mean PI fluorescence intensity. Sel1L −/− ( 1-3 ) represent three independent HepG2 cell clones. D–F, MTT assays of Sel1L +/+ and Sel1L −/− cells after treatment with increased concentrations of TNF-α ( D ), IL-12 ( E ), and TNFα + IL-12 ( F ). G and H, fluorescence reporter assay of ER stress in TNF-α ( G ) and IL-12 ( H ) treated Sel1L +/+ and Sel1L −/− cells. I, MTT assay of Sel1L +/+ and Sel1L −/− cells after treatment with TNFα or TNFα + Nec-1. J , FITC and PE dual channel FACS assay of Sel1L +/+ and Sel1L −/− cells after treatment with PBS (Mock), TNF-α, TNF-α + Ru360, or TNF-α + NAC. All data are mean ± S.E. ( n = 3). *, p < 0.05; **, p < 0.01 by Student's t test.
    Optimized Crispr Design Tool, supplied by Synthego Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/optimized crispr design tool/product/Synthego Inc
    Average 90 stars, based on 1 article reviews
    optimized crispr design tool - by Bioz Stars, 2026-03
    90/100 stars

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    1) Product Images from "ERAD deficiency promotes mitochondrial dysfunction and transcriptional rewiring in human hepatic cells"

    Article Title: ERAD deficiency promotes mitochondrial dysfunction and transcriptional rewiring in human hepatic cells

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.RA120.013987

    Genetic ablation of ERAD sensitizes HepG2 cells to TNFα– and IL-12–induced cell death. A, diagram illustrating CRISPR/Cas9-mediated editing of the Sel1L gene in HepG2 cells. F, R1, and R2 represent PCR primers used for genotyping of Sel1L-edited cells. Black bars represent exons. B , top , PCR analysis of Sel1L-normal (Sel1L +/+ ) and deleted (Sel1L −/− ) cells. Bottom, Western blotting analysis of SEL1L expression in Sel1L +/+ and Sel1L −/− cells; the upper band is a nonspecific band. C, average cell death level of Sel1L +/+ and Sel1L −/− cells as indicated by the mean PI fluorescence intensity. Sel1L −/− ( 1-3 ) represent three independent HepG2 cell clones. D–F, MTT assays of Sel1L +/+ and Sel1L −/− cells after treatment with increased concentrations of TNF-α ( D ), IL-12 ( E ), and TNFα + IL-12 ( F ). G and H, fluorescence reporter assay of ER stress in TNF-α ( G ) and IL-12 ( H ) treated Sel1L +/+ and Sel1L −/− cells. I, MTT assay of Sel1L +/+ and Sel1L −/− cells after treatment with TNFα or TNFα + Nec-1. J , FITC and PE dual channel FACS assay of Sel1L +/+ and Sel1L −/− cells after treatment with PBS (Mock), TNF-α, TNF-α + Ru360, or TNF-α + NAC. All data are mean ± S.E. ( n = 3). *, p < 0.05; **, p < 0.01 by Student's t test.
    Figure Legend Snippet: Genetic ablation of ERAD sensitizes HepG2 cells to TNFα– and IL-12–induced cell death. A, diagram illustrating CRISPR/Cas9-mediated editing of the Sel1L gene in HepG2 cells. F, R1, and R2 represent PCR primers used for genotyping of Sel1L-edited cells. Black bars represent exons. B , top , PCR analysis of Sel1L-normal (Sel1L +/+ ) and deleted (Sel1L −/− ) cells. Bottom, Western blotting analysis of SEL1L expression in Sel1L +/+ and Sel1L −/− cells; the upper band is a nonspecific band. C, average cell death level of Sel1L +/+ and Sel1L −/− cells as indicated by the mean PI fluorescence intensity. Sel1L −/− ( 1-3 ) represent three independent HepG2 cell clones. D–F, MTT assays of Sel1L +/+ and Sel1L −/− cells after treatment with increased concentrations of TNF-α ( D ), IL-12 ( E ), and TNFα + IL-12 ( F ). G and H, fluorescence reporter assay of ER stress in TNF-α ( G ) and IL-12 ( H ) treated Sel1L +/+ and Sel1L −/− cells. I, MTT assay of Sel1L +/+ and Sel1L −/− cells after treatment with TNFα or TNFα + Nec-1. J , FITC and PE dual channel FACS assay of Sel1L +/+ and Sel1L −/− cells after treatment with PBS (Mock), TNF-α, TNF-α + Ru360, or TNF-α + NAC. All data are mean ± S.E. ( n = 3). *, p < 0.05; **, p < 0.01 by Student's t test.

    Techniques Used: CRISPR, Western Blot, Expressing, Fluorescence, Clone Assay, Reporter Assay, MTT Assay



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    Genetic ablation of ERAD sensitizes HepG2 cells to TNFα– and IL-12–induced cell death. A, diagram <t>illustrating</t> <t>CRISPR/Cas9-mediated</t> editing of the Sel1L gene in HepG2 cells. F, R1, and R2 represent PCR primers used for genotyping of Sel1L-edited cells. Black bars represent exons. B , top , PCR analysis of Sel1L-normal (Sel1L +/+ ) and deleted (Sel1L −/− ) cells. Bottom, Western blotting analysis of SEL1L expression in Sel1L +/+ and Sel1L −/− cells; the upper band is a nonspecific band. C, average cell death level of Sel1L +/+ and Sel1L −/− cells as indicated by the mean PI fluorescence intensity. Sel1L −/− ( 1-3 ) represent three independent HepG2 cell clones. D–F, MTT assays of Sel1L +/+ and Sel1L −/− cells after treatment with increased concentrations of TNF-α ( D ), IL-12 ( E ), and TNFα + IL-12 ( F ). G and H, fluorescence reporter assay of ER stress in TNF-α ( G ) and IL-12 ( H ) treated Sel1L +/+ and Sel1L −/− cells. I, MTT assay of Sel1L +/+ and Sel1L −/− cells after treatment with TNFα or TNFα + Nec-1. J , FITC and PE dual channel FACS assay of Sel1L +/+ and Sel1L −/− cells after treatment with PBS (Mock), TNF-α, TNF-α + Ru360, or TNF-α + NAC. All data are mean ± S.E. ( n = 3). *, p < 0.05; **, p < 0.01 by Student's t test.
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    Genetic ablation of ERAD sensitizes HepG2 cells to TNFα– and IL-12–induced cell death. A, diagram <t>illustrating</t> <t>CRISPR/Cas9-mediated</t> editing of the Sel1L gene in HepG2 cells. F, R1, and R2 represent PCR primers used for genotyping of Sel1L-edited cells. Black bars represent exons. B , top , PCR analysis of Sel1L-normal (Sel1L +/+ ) and deleted (Sel1L −/− ) cells. Bottom, Western blotting analysis of SEL1L expression in Sel1L +/+ and Sel1L −/− cells; the upper band is a nonspecific band. C, average cell death level of Sel1L +/+ and Sel1L −/− cells as indicated by the mean PI fluorescence intensity. Sel1L −/− ( 1-3 ) represent three independent HepG2 cell clones. D–F, MTT assays of Sel1L +/+ and Sel1L −/− cells after treatment with increased concentrations of TNF-α ( D ), IL-12 ( E ), and TNFα + IL-12 ( F ). G and H, fluorescence reporter assay of ER stress in TNF-α ( G ) and IL-12 ( H ) treated Sel1L +/+ and Sel1L −/− cells. I, MTT assay of Sel1L +/+ and Sel1L −/− cells after treatment with TNFα or TNFα + Nec-1. J , FITC and PE dual channel FACS assay of Sel1L +/+ and Sel1L −/− cells after treatment with PBS (Mock), TNF-α, TNF-α + Ru360, or TNF-α + NAC. All data are mean ± S.E. ( n = 3). *, p < 0.05; **, p < 0.01 by Student's t test.
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    Image Search Results


    Genetic ablation of ERAD sensitizes HepG2 cells to TNFα– and IL-12–induced cell death. A, diagram illustrating CRISPR/Cas9-mediated editing of the Sel1L gene in HepG2 cells. F, R1, and R2 represent PCR primers used for genotyping of Sel1L-edited cells. Black bars represent exons. B , top , PCR analysis of Sel1L-normal (Sel1L +/+ ) and deleted (Sel1L −/− ) cells. Bottom, Western blotting analysis of SEL1L expression in Sel1L +/+ and Sel1L −/− cells; the upper band is a nonspecific band. C, average cell death level of Sel1L +/+ and Sel1L −/− cells as indicated by the mean PI fluorescence intensity. Sel1L −/− ( 1-3 ) represent three independent HepG2 cell clones. D–F, MTT assays of Sel1L +/+ and Sel1L −/− cells after treatment with increased concentrations of TNF-α ( D ), IL-12 ( E ), and TNFα + IL-12 ( F ). G and H, fluorescence reporter assay of ER stress in TNF-α ( G ) and IL-12 ( H ) treated Sel1L +/+ and Sel1L −/− cells. I, MTT assay of Sel1L +/+ and Sel1L −/− cells after treatment with TNFα or TNFα + Nec-1. J , FITC and PE dual channel FACS assay of Sel1L +/+ and Sel1L −/− cells after treatment with PBS (Mock), TNF-α, TNF-α + Ru360, or TNF-α + NAC. All data are mean ± S.E. ( n = 3). *, p < 0.05; **, p < 0.01 by Student's t test.

    Journal: The Journal of Biological Chemistry

    Article Title: ERAD deficiency promotes mitochondrial dysfunction and transcriptional rewiring in human hepatic cells

    doi: 10.1074/jbc.RA120.013987

    Figure Lengend Snippet: Genetic ablation of ERAD sensitizes HepG2 cells to TNFα– and IL-12–induced cell death. A, diagram illustrating CRISPR/Cas9-mediated editing of the Sel1L gene in HepG2 cells. F, R1, and R2 represent PCR primers used for genotyping of Sel1L-edited cells. Black bars represent exons. B , top , PCR analysis of Sel1L-normal (Sel1L +/+ ) and deleted (Sel1L −/− ) cells. Bottom, Western blotting analysis of SEL1L expression in Sel1L +/+ and Sel1L −/− cells; the upper band is a nonspecific band. C, average cell death level of Sel1L +/+ and Sel1L −/− cells as indicated by the mean PI fluorescence intensity. Sel1L −/− ( 1-3 ) represent three independent HepG2 cell clones. D–F, MTT assays of Sel1L +/+ and Sel1L −/− cells after treatment with increased concentrations of TNF-α ( D ), IL-12 ( E ), and TNFα + IL-12 ( F ). G and H, fluorescence reporter assay of ER stress in TNF-α ( G ) and IL-12 ( H ) treated Sel1L +/+ and Sel1L −/− cells. I, MTT assay of Sel1L +/+ and Sel1L −/− cells after treatment with TNFα or TNFα + Nec-1. J , FITC and PE dual channel FACS assay of Sel1L +/+ and Sel1L −/− cells after treatment with PBS (Mock), TNF-α, TNF-α + Ru360, or TNF-α + NAC. All data are mean ± S.E. ( n = 3). *, p < 0.05; **, p < 0.01 by Student's t test.

    Article Snippet: For Cas9-mediated gene editing, sgRNAs flanking exon 6 were designed using the Optimized CRISPR Design tool ( https://www.synthego.com/products/bioinformatics/crispr-design-tool ) and cloned into the expression vector pGL3-U6-2sgRNA.

    Techniques: CRISPR, Western Blot, Expressing, Fluorescence, Clone Assay, Reporter Assay, MTT Assay