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flp recombinase expression cassette  (Addgene inc)


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    Structured Review

    Addgene inc flp recombinase expression cassette
    (A) Mutant mESC lines were generated by <t>recombinase-mediated</t> cassette exchange (RMCE). Heterotypic Flp recognition target (FRT) sites and an mCherry expression reporter were inserted around Klf4 enhancer E2 in mESCs. In the presence of an exchange plasmid, which contains the enhancer E2 flanked by the heterotypic FRT sites, Flp recombinase facilitates integration of the desired enhancer E2 sequence. Single cells were selected after RMCE, isolated for clonal outgrowth, and confirmed by PCR and sequencing. (B-F) Analysis of gene expression in the mutant mESC lines. For each enhancer E2 sequence, multiple clonal populations were grown under naïve-state conditions, and total mRNA was harvested from each and reverse transcribed into cDNA. Quantitative PCR was performed on the cDNA to measure levels of Klf4 (B), Rad23b (C), Pou5f1 (D), Sox2 (E), Nanog (F), Esrrb (G), Tbx3 (H), Klf2 (I), and Klf5 (J). Data was analyzed using the ΔΔCT method. Each data point represents the average from 2–3 technical replicates for each isolated clone, and error bars represent mean of clones ± 95% confidence interval. Asterisks represent statistically significant differences between averages (*p<0.05; **p<0.01, unpaired t-test), with brackets to indicate samples compared. If not shown, averages were not statistically different. Outliers, as determined by the Grubb’s test, are denoted by unfilled shapes and were not utilized in statistical tests.
    Flp Recombinase Expression Cassette, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flp recombinase expression cassette/product/Addgene inc
    Average 92 stars, based on 21 article reviews
    flp recombinase expression cassette - by Bioz Stars, 2026-02
    92/100 stars

    Images

    1) Product Images from "A suboptimal OCT4-SOX2 binding site facilitates the naïve-state specific function of a Klf4 enhancer"

    Article Title: A suboptimal OCT4-SOX2 binding site facilitates the naïve-state specific function of a Klf4 enhancer

    Journal: PLOS ONE

    doi: 10.1371/journal.pone.0311120

    (A) Mutant mESC lines were generated by recombinase-mediated cassette exchange (RMCE). Heterotypic Flp recognition target (FRT) sites and an mCherry expression reporter were inserted around Klf4 enhancer E2 in mESCs. In the presence of an exchange plasmid, which contains the enhancer E2 flanked by the heterotypic FRT sites, Flp recombinase facilitates integration of the desired enhancer E2 sequence. Single cells were selected after RMCE, isolated for clonal outgrowth, and confirmed by PCR and sequencing. (B-F) Analysis of gene expression in the mutant mESC lines. For each enhancer E2 sequence, multiple clonal populations were grown under naïve-state conditions, and total mRNA was harvested from each and reverse transcribed into cDNA. Quantitative PCR was performed on the cDNA to measure levels of Klf4 (B), Rad23b (C), Pou5f1 (D), Sox2 (E), Nanog (F), Esrrb (G), Tbx3 (H), Klf2 (I), and Klf5 (J). Data was analyzed using the ΔΔCT method. Each data point represents the average from 2–3 technical replicates for each isolated clone, and error bars represent mean of clones ± 95% confidence interval. Asterisks represent statistically significant differences between averages (*p<0.05; **p<0.01, unpaired t-test), with brackets to indicate samples compared. If not shown, averages were not statistically different. Outliers, as determined by the Grubb’s test, are denoted by unfilled shapes and were not utilized in statistical tests.
    Figure Legend Snippet: (A) Mutant mESC lines were generated by recombinase-mediated cassette exchange (RMCE). Heterotypic Flp recognition target (FRT) sites and an mCherry expression reporter were inserted around Klf4 enhancer E2 in mESCs. In the presence of an exchange plasmid, which contains the enhancer E2 flanked by the heterotypic FRT sites, Flp recombinase facilitates integration of the desired enhancer E2 sequence. Single cells were selected after RMCE, isolated for clonal outgrowth, and confirmed by PCR and sequencing. (B-F) Analysis of gene expression in the mutant mESC lines. For each enhancer E2 sequence, multiple clonal populations were grown under naïve-state conditions, and total mRNA was harvested from each and reverse transcribed into cDNA. Quantitative PCR was performed on the cDNA to measure levels of Klf4 (B), Rad23b (C), Pou5f1 (D), Sox2 (E), Nanog (F), Esrrb (G), Tbx3 (H), Klf2 (I), and Klf5 (J). Data was analyzed using the ΔΔCT method. Each data point represents the average from 2–3 technical replicates for each isolated clone, and error bars represent mean of clones ± 95% confidence interval. Asterisks represent statistically significant differences between averages (*p<0.05; **p<0.01, unpaired t-test), with brackets to indicate samples compared. If not shown, averages were not statistically different. Outliers, as determined by the Grubb’s test, are denoted by unfilled shapes and were not utilized in statistical tests.

    Techniques Used: Mutagenesis, Generated, Expressing, Plasmid Preparation, Sequencing, Isolation, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Clone Assay



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    flp recombinase expression cassette  (Addgene inc)
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    Addgene inc flp recombinase expression cassette
    (A) Mutant mESC lines were generated by <t>recombinase-mediated</t> cassette exchange (RMCE). Heterotypic Flp recognition target (FRT) sites and an mCherry expression reporter were inserted around Klf4 enhancer E2 in mESCs. In the presence of an exchange plasmid, which contains the enhancer E2 flanked by the heterotypic FRT sites, Flp recombinase facilitates integration of the desired enhancer E2 sequence. Single cells were selected after RMCE, isolated for clonal outgrowth, and confirmed by PCR and sequencing. (B-F) Analysis of gene expression in the mutant mESC lines. For each enhancer E2 sequence, multiple clonal populations were grown under naïve-state conditions, and total mRNA was harvested from each and reverse transcribed into cDNA. Quantitative PCR was performed on the cDNA to measure levels of Klf4 (B), Rad23b (C), Pou5f1 (D), Sox2 (E), Nanog (F), Esrrb (G), Tbx3 (H), Klf2 (I), and Klf5 (J). Data was analyzed using the ΔΔCT method. Each data point represents the average from 2–3 technical replicates for each isolated clone, and error bars represent mean of clones ± 95% confidence interval. Asterisks represent statistically significant differences between averages (*p<0.05; **p<0.01, unpaired t-test), with brackets to indicate samples compared. If not shown, averages were not statistically different. Outliers, as determined by the Grubb’s test, are denoted by unfilled shapes and were not utilized in statistical tests.
    Flp Recombinase Expression Cassette, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/flp recombinase expression cassette/product/Addgene inc
    Average 92 stars, based on 1 article reviews
    flp recombinase expression cassette - by Bioz Stars, 2026-02
    92/100 stars
    		  Buy from Supplier
    flp recombinase expression cassettes  (GenScript corporation)
    90
    GenScript corporation flp recombinase expression cassettes
    (A) Mutant mESC lines were generated by <t>recombinase-mediated</t> cassette exchange (RMCE). Heterotypic Flp recognition target (FRT) sites and an mCherry expression reporter were inserted around Klf4 enhancer E2 in mESCs. In the presence of an exchange plasmid, which contains the enhancer E2 flanked by the heterotypic FRT sites, Flp recombinase facilitates integration of the desired enhancer E2 sequence. Single cells were selected after RMCE, isolated for clonal outgrowth, and confirmed by PCR and sequencing. (B-F) Analysis of gene expression in the mutant mESC lines. For each enhancer E2 sequence, multiple clonal populations were grown under naïve-state conditions, and total mRNA was harvested from each and reverse transcribed into cDNA. Quantitative PCR was performed on the cDNA to measure levels of Klf4 (B), Rad23b (C), Pou5f1 (D), Sox2 (E), Nanog (F), Esrrb (G), Tbx3 (H), Klf2 (I), and Klf5 (J). Data was analyzed using the ΔΔCT method. Each data point represents the average from 2–3 technical replicates for each isolated clone, and error bars represent mean of clones ± 95% confidence interval. Asterisks represent statistically significant differences between averages (*p<0.05; **p<0.01, unpaired t-test), with brackets to indicate samples compared. If not shown, averages were not statistically different. Outliers, as determined by the Grubb’s test, are denoted by unfilled shapes and were not utilized in statistical tests.
    Flp Recombinase Expression Cassettes, supplied by GenScript corporation, 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/flp recombinase expression cassettes/product/GenScript corporation
    Average 90 stars, based on 1 article reviews
    flp recombinase expression cassettes - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    (A) Mutant mESC lines were generated by recombinase-mediated cassette exchange (RMCE). Heterotypic Flp recognition target (FRT) sites and an mCherry expression reporter were inserted around Klf4 enhancer E2 in mESCs. In the presence of an exchange plasmid, which contains the enhancer E2 flanked by the heterotypic FRT sites, Flp recombinase facilitates integration of the desired enhancer E2 sequence. Single cells were selected after RMCE, isolated for clonal outgrowth, and confirmed by PCR and sequencing. (B-F) Analysis of gene expression in the mutant mESC lines. For each enhancer E2 sequence, multiple clonal populations were grown under naïve-state conditions, and total mRNA was harvested from each and reverse transcribed into cDNA. Quantitative PCR was performed on the cDNA to measure levels of Klf4 (B), Rad23b (C), Pou5f1 (D), Sox2 (E), Nanog (F), Esrrb (G), Tbx3 (H), Klf2 (I), and Klf5 (J). Data was analyzed using the ΔΔCT method. Each data point represents the average from 2–3 technical replicates for each isolated clone, and error bars represent mean of clones ± 95% confidence interval. Asterisks represent statistically significant differences between averages (*p<0.05; **p<0.01, unpaired t-test), with brackets to indicate samples compared. If not shown, averages were not statistically different. Outliers, as determined by the Grubb’s test, are denoted by unfilled shapes and were not utilized in statistical tests.

    Journal: PLOS ONE

    Article Title: A suboptimal OCT4-SOX2 binding site facilitates the naïve-state specific function of a Klf4 enhancer

    doi: 10.1371/journal.pone.0311120

    Figure Lengend Snippet: (A) Mutant mESC lines were generated by recombinase-mediated cassette exchange (RMCE). Heterotypic Flp recognition target (FRT) sites and an mCherry expression reporter were inserted around Klf4 enhancer E2 in mESCs. In the presence of an exchange plasmid, which contains the enhancer E2 flanked by the heterotypic FRT sites, Flp recombinase facilitates integration of the desired enhancer E2 sequence. Single cells were selected after RMCE, isolated for clonal outgrowth, and confirmed by PCR and sequencing. (B-F) Analysis of gene expression in the mutant mESC lines. For each enhancer E2 sequence, multiple clonal populations were grown under naïve-state conditions, and total mRNA was harvested from each and reverse transcribed into cDNA. Quantitative PCR was performed on the cDNA to measure levels of Klf4 (B), Rad23b (C), Pou5f1 (D), Sox2 (E), Nanog (F), Esrrb (G), Tbx3 (H), Klf2 (I), and Klf5 (J). Data was analyzed using the ΔΔCT method. Each data point represents the average from 2–3 technical replicates for each isolated clone, and error bars represent mean of clones ± 95% confidence interval. Asterisks represent statistically significant differences between averages (*p<0.05; **p<0.01, unpaired t-test), with brackets to indicate samples compared. If not shown, averages were not statistically different. Outliers, as determined by the Grubb’s test, are denoted by unfilled shapes and were not utilized in statistical tests.

    Article Snippet: The PGK promoter was inserted at the KpnI/XhoI cut sites; the optimized Flp recombinase expression cassette was amplified from pDIRE (Addgene #26745) and inserted at EcoRI/NotI; and a cassette with an internal ribosome entry sequence (IRES) sequence followed by the puromycin resistance gene was inserted at NotI/SacI.

    Techniques: Mutagenesis, Generated, Expressing, Plasmid Preparation, Sequencing, Isolation, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction, Clone Assay