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i scei expression plasmid  (Addgene inc)


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    Addgene inc i scei expression plasmid
    I Scei Expression Plasmid, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 274 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/i+scei+plasmid/bio_rxiv__64898__2026__02__17__706442-228-11-14?v=Addgene+inc
    Average 96 stars, based on 274 article reviews
    i scei expression plasmid - by Bioz Stars, 2026-07
    96/100 stars

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    (A, D) Schematics of the HR-EGFP/5′EGFP (A) and PIMEJ5GFP (D) reporter constructs. HR-EGFP/5′EGFP includes a mutated EGFP gene containing an I-SceI recognition site and a truncated 5′EGFP sequence serving as a repair template. PIMEJ5GFP contains two distal I-SceI sites flanking a puromycin gene and the EGFP sequence. Stable U2OS clones with chromosomally integrated reporters were transfected with siCTRL, siBAP1, or siBRCA1. The following day, cells were <t>transfected</t> <t>with</t> <t>I-SceI-RFP</t> (red fluorescence) and treated with triamcinolone acetonide (TCA) 24 h later to induce nuclear translocation of I-SceI. RFP+ cells were analyzed for GFP fluorescence. (B, E) Representative flow cytometry plots showing GFP+ cells (yellow gate) within the RFP+ population under each condition. GFP fluorescence: B525-FITC-A; side scatter: SSC. (C, F) Bar plots quantifying HR (C) and NHEJ (F) repair efficiency across ≥3 independent experiments (dots). Data normalized to control and presented as mean ± SD. ****p < 0.0001. P-values by one-way ANOVA with Tukey’s correction (C) or paired t-test (F).
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    (A, D) Schematics of the HR-EGFP/5′EGFP (A) and PIMEJ5GFP (D) reporter constructs. HR-EGFP/5′EGFP includes a mutated EGFP gene containing an I-SceI recognition site and a truncated 5′EGFP sequence serving as a repair template. PIMEJ5GFP contains two distal I-SceI sites flanking a puromycin gene and the EGFP sequence. Stable U2OS clones with chromosomally integrated reporters were transfected with siCTRL, siBAP1, or siBRCA1. The following day, cells were <t>transfected</t> <t>with</t> <t>I-SceI-RFP</t> (red fluorescence) and treated with triamcinolone acetonide (TCA) 24 h later to induce nuclear translocation of I-SceI. RFP+ cells were analyzed for GFP fluorescence. (B, E) Representative flow cytometry plots showing GFP+ cells (yellow gate) within the RFP+ population under each condition. GFP fluorescence: B525-FITC-A; side scatter: SSC. (C, F) Bar plots quantifying HR (C) and NHEJ (F) repair efficiency across ≥3 independent experiments (dots). Data normalized to control and presented as mean ± SD. ****p < 0.0001. P-values by one-way ANOVA with Tukey’s correction (C) or paired t-test (F).
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    (A, D) Schematics of the HR-EGFP/5′EGFP (A) and PIMEJ5GFP (D) reporter constructs. HR-EGFP/5′EGFP includes a mutated EGFP gene containing an I-SceI recognition site and a truncated 5′EGFP sequence serving as a repair template. PIMEJ5GFP contains two distal I-SceI sites flanking a puromycin gene and the EGFP sequence. Stable U2OS clones with chromosomally integrated reporters were transfected with siCTRL, siBAP1, or siBRCA1. The following day, cells were transfected with I-SceI-RFP (red fluorescence) and treated with triamcinolone acetonide (TCA) 24 h later to induce nuclear translocation of I-SceI. RFP+ cells were analyzed for GFP fluorescence. (B, E) Representative flow cytometry plots showing GFP+ cells (yellow gate) within the RFP+ population under each condition. GFP fluorescence: B525-FITC-A; side scatter: SSC. (C, F) Bar plots quantifying HR (C) and NHEJ (F) repair efficiency across ≥3 independent experiments (dots). Data normalized to control and presented as mean ± SD. ****p < 0.0001. P-values by one-way ANOVA with Tukey’s correction (C) or paired t-test (F).

    Journal: bioRxiv

    Article Title: BAP1 loss impairs Non-Homologous End Joining DNA repair promoting genomic instability

    doi: 10.64898/2026.02.17.706330

    Figure Lengend Snippet: (A, D) Schematics of the HR-EGFP/5′EGFP (A) and PIMEJ5GFP (D) reporter constructs. HR-EGFP/5′EGFP includes a mutated EGFP gene containing an I-SceI recognition site and a truncated 5′EGFP sequence serving as a repair template. PIMEJ5GFP contains two distal I-SceI sites flanking a puromycin gene and the EGFP sequence. Stable U2OS clones with chromosomally integrated reporters were transfected with siCTRL, siBAP1, or siBRCA1. The following day, cells were transfected with I-SceI-RFP (red fluorescence) and treated with triamcinolone acetonide (TCA) 24 h later to induce nuclear translocation of I-SceI. RFP+ cells were analyzed for GFP fluorescence. (B, E) Representative flow cytometry plots showing GFP+ cells (yellow gate) within the RFP+ population under each condition. GFP fluorescence: B525-FITC-A; side scatter: SSC. (C, F) Bar plots quantifying HR (C) and NHEJ (F) repair efficiency across ≥3 independent experiments (dots). Data normalized to control and presented as mean ± SD. ****p < 0.0001. P-values by one-way ANOVA with Tukey’s correction (C) or paired t-test (F).

    Article Snippet: All U2OS clones were transfected with 2 μg I-SceI-RFP plasmid (Addgene, #17654) and after 24 hours, treated with 0.4 μM triamcinolone acetonide (TCA) or DMSO.

    Techniques: Construct, Sequencing, Clone Assay, Transfection, Fluorescence, Translocation Assay, Flow Cytometry, Control