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tale tool box kit  (Addgene inc)


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    Structured Review

    Addgene inc tale tool box kit
    (A) 500kb of linear genome (mm9, chromosome 6:135,623,529-136,123,529), <t>encompassing</t> <t>Grin2b,</t> including TSS (green), proximal intronic sequences targeted by SETDB1 (yellow), and two loop loopings (red) (1) Grin2bTSS+378kb and (2) Grin2bTSS+471kb, with sequences homologue to human GRIN2BTSS+348kb and GRIN2BTSS+449kb (Fig. S1) (B) (Top) Browser tracks for histone marks H3K4me3 and H3K27ac in adult mouse cerebral cortex(Dixon et al., 2012). (C) Fold-change (CK-Setdb1 transgenic (Tg) /wildtype littermate (Wt) of 3C PCR from adult Tg and Wt cortex (see also Fig. S2). Notice increased physical interactions of (yellow) TSS-bound +15 to +40kb intronic sequence, and significant decrease in (red box no. 1) Grin2bTSS+378kb and (red box no. 2) Grin2bTSS+471kb. Two-way ANOVA, 3C interaction × genotype F(7,32)=54.905(p<0.001). Newman-Keuls post-hoc *, **; P< 0.05, 0.01. (D) Quantification of Grin2b (left) RNA and (right) protein in adult (6–8 week) CK-Setdb1 Tg and wildtype (Wt) littermate control cortex. mean ± S.D, N=5 (RNA) and N=3 (immunoblot)/group. (E) Activity-dependent regulation of Grin2b higher order chromatin in hippocampal neurons. (top, left to right) Grin2b RNA, 3C quantification of Grin2bTSS+378kb and Grin2bTSS+471kb and SETDB1 occupancy across four regulatory sequences at Grin2b locus. Note significant increase at SETDB1 target site after 15 hours of picrotoxin (PTX) or vehicle control (DMSO). N=3–6 experiments/group, data shown as mean ± S.D..(F) (left) <t>Grin2b-TALE-VP64-GFP</t> specifically targets mouse Grin2b loop no. 2 (Grin2bTSS+471kb), 471 kb downstream of TSS. Images show Neuro2A cells and cultured hippocampal neurons expressing GFP-tagged TALE-VP64. RT-PCR for Grin2b and Gapdh control showing specific expression in cultured neurons. (right) anti-TALE-VP64 ChIP in N2A and NG108 cells and primary cortical neurons, expressed (y-axis) as fold change compared to non-transfected condition (N=3/group; (mean ± S.E.M., * Two-way ANOVA cell type × TALE-VP64 binding F(2,12)=4.04, P<0.05, Bonferroni posthoc P < 0.05).RT-PCR Grin2b, Grin2a and Setdb1 RNA levels in neurons transfected with TALE-VP64-EGFP (TALE), compared to mock-transfected neurons (Con). Notice specific TALE-VP64 mediated increase in Grin2b RNA (N = 6 per group, mean ± S.E.M, *P < 0.05, t-test). See also Figures S1 and S2.
    Tale Tool Box Kit, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cell+transcriptomic+sequencing+dataset/TALE+Toolbox+(Kit+%231000000019)/pmc04258154-224-49-53
    Average 93 stars, based on 25 article reviews
    tale tool box kit - by Bioz Stars, 2026-11
    93/100 stars

    Images

    1) Product Images from "CONSERVED HIGHER ORDER CHROMATIN REGULATES NMDA RECEPTOR GENE EXPRESSION AND COGNITION"

    Article Title: CONSERVED HIGHER ORDER CHROMATIN REGULATES NMDA RECEPTOR GENE EXPRESSION AND COGNITION

    Journal: Neuron

    doi: 10.1016/j.neuron.2014.10.032

    (A) 500kb of linear genome (mm9, chromosome 6:135,623,529-136,123,529), encompassing Grin2b, including TSS (green), proximal intronic sequences targeted by SETDB1 (yellow), and two loop loopings (red) (1) Grin2bTSS+378kb and (2) Grin2bTSS+471kb, with sequences homologue to human GRIN2BTSS+348kb and GRIN2BTSS+449kb (Fig. S1) (B) (Top) Browser tracks for histone marks H3K4me3 and H3K27ac in adult mouse cerebral cortex(Dixon et al., 2012). (C) Fold-change (CK-Setdb1 transgenic (Tg) /wildtype littermate (Wt) of 3C PCR from adult Tg and Wt cortex (see also Fig. S2). Notice increased physical interactions of (yellow) TSS-bound +15 to +40kb intronic sequence, and significant decrease in (red box no. 1) Grin2bTSS+378kb and (red box no. 2) Grin2bTSS+471kb. Two-way ANOVA, 3C interaction × genotype F(7,32)=54.905(p<0.001). Newman-Keuls post-hoc *, **; P< 0.05, 0.01. (D) Quantification of Grin2b (left) RNA and (right) protein in adult (6–8 week) CK-Setdb1 Tg and wildtype (Wt) littermate control cortex. mean ± S.D, N=5 (RNA) and N=3 (immunoblot)/group. (E) Activity-dependent regulation of Grin2b higher order chromatin in hippocampal neurons. (top, left to right) Grin2b RNA, 3C quantification of Grin2bTSS+378kb and Grin2bTSS+471kb and SETDB1 occupancy across four regulatory sequences at Grin2b locus. Note significant increase at SETDB1 target site after 15 hours of picrotoxin (PTX) or vehicle control (DMSO). N=3–6 experiments/group, data shown as mean ± S.D..(F) (left) Grin2b-TALE-VP64-GFP specifically targets mouse Grin2b loop no. 2 (Grin2bTSS+471kb), 471 kb downstream of TSS. Images show Neuro2A cells and cultured hippocampal neurons expressing GFP-tagged TALE-VP64. RT-PCR for Grin2b and Gapdh control showing specific expression in cultured neurons. (right) anti-TALE-VP64 ChIP in N2A and NG108 cells and primary cortical neurons, expressed (y-axis) as fold change compared to non-transfected condition (N=3/group; (mean ± S.E.M., * Two-way ANOVA cell type × TALE-VP64 binding F(2,12)=4.04, P<0.05, Bonferroni posthoc P < 0.05).RT-PCR Grin2b, Grin2a and Setdb1 RNA levels in neurons transfected with TALE-VP64-EGFP (TALE), compared to mock-transfected neurons (Con). Notice specific TALE-VP64 mediated increase in Grin2b RNA (N = 6 per group, mean ± S.E.M, *P < 0.05, t-test). See also Figures S1 and S2.
    Figure Legend Snippet: (A) 500kb of linear genome (mm9, chromosome 6:135,623,529-136,123,529), encompassing Grin2b, including TSS (green), proximal intronic sequences targeted by SETDB1 (yellow), and two loop loopings (red) (1) Grin2bTSS+378kb and (2) Grin2bTSS+471kb, with sequences homologue to human GRIN2BTSS+348kb and GRIN2BTSS+449kb (Fig. S1) (B) (Top) Browser tracks for histone marks H3K4me3 and H3K27ac in adult mouse cerebral cortex(Dixon et al., 2012). (C) Fold-change (CK-Setdb1 transgenic (Tg) /wildtype littermate (Wt) of 3C PCR from adult Tg and Wt cortex (see also Fig. S2). Notice increased physical interactions of (yellow) TSS-bound +15 to +40kb intronic sequence, and significant decrease in (red box no. 1) Grin2bTSS+378kb and (red box no. 2) Grin2bTSS+471kb. Two-way ANOVA, 3C interaction × genotype F(7,32)=54.905(p<0.001). Newman-Keuls post-hoc *, **; P< 0.05, 0.01. (D) Quantification of Grin2b (left) RNA and (right) protein in adult (6–8 week) CK-Setdb1 Tg and wildtype (Wt) littermate control cortex. mean ± S.D, N=5 (RNA) and N=3 (immunoblot)/group. (E) Activity-dependent regulation of Grin2b higher order chromatin in hippocampal neurons. (top, left to right) Grin2b RNA, 3C quantification of Grin2bTSS+378kb and Grin2bTSS+471kb and SETDB1 occupancy across four regulatory sequences at Grin2b locus. Note significant increase at SETDB1 target site after 15 hours of picrotoxin (PTX) or vehicle control (DMSO). N=3–6 experiments/group, data shown as mean ± S.D..(F) (left) Grin2b-TALE-VP64-GFP specifically targets mouse Grin2b loop no. 2 (Grin2bTSS+471kb), 471 kb downstream of TSS. Images show Neuro2A cells and cultured hippocampal neurons expressing GFP-tagged TALE-VP64. RT-PCR for Grin2b and Gapdh control showing specific expression in cultured neurons. (right) anti-TALE-VP64 ChIP in N2A and NG108 cells and primary cortical neurons, expressed (y-axis) as fold change compared to non-transfected condition (N=3/group; (mean ± S.E.M., * Two-way ANOVA cell type × TALE-VP64 binding F(2,12)=4.04, P<0.05, Bonferroni posthoc P < 0.05).RT-PCR Grin2b, Grin2a and Setdb1 RNA levels in neurons transfected with TALE-VP64-EGFP (TALE), compared to mock-transfected neurons (Con). Notice specific TALE-VP64 mediated increase in Grin2b RNA (N = 6 per group, mean ± S.E.M, *P < 0.05, t-test). See also Figures S1 and S2.

    Techniques Used: Transgenic Assay, Sequencing, Control, Western Blot, Activity Assay, Cell Culture, Expressing, Reverse Transcription Polymerase Chain Reaction, Transfection, Binding Assay

    Related Articles

    Modification:

    Article Title: Tet1 Isoforms Differentially Regulate Gene Expression, Synaptic Transmission, and Memory in the Mammalian Brain
    Article Snippet: Briefly, primers were designed using NEBuilder Assembly Tool (v1.12.18) to generate overlapping (20–25 bp) PCR fragments amplified using Q5 High-Fidelity DNA Polymerase (NEB). .. Modified TALE constructs were created using PCR fragments amplified from pAAV-CW3SL-EGFP (Addgene #61463), a gift from Bong-Kiun Kaang , and constructs contained in the TALE Toolbox (Addgene kit #1000000019), a gift from Feng Zhang. ..

    Construct:

    Article Title: Tet1 Isoforms Differentially Regulate Gene Expression, Synaptic Transmission, and Memory in the Mammalian Brain
    Article Snippet: Briefly, primers were designed using NEBuilder Assembly Tool (v1.12.18) to generate overlapping (20–25 bp) PCR fragments amplified using Q5 High-Fidelity DNA Polymerase (NEB). .. Modified TALE constructs were created using PCR fragments amplified from pAAV-CW3SL-EGFP (Addgene #61463), a gift from Bong-Kiun Kaang , and constructs contained in the TALE Toolbox (Addgene kit #1000000019), a gift from Feng Zhang. ..

    Polymerase Chain Reaction:

    Article Title: Tet1 Isoforms Differentially Regulate Gene Expression, Synaptic Transmission, and Memory in the Mammalian Brain
    Article Snippet: Briefly, primers were designed using NEBuilder Assembly Tool (v1.12.18) to generate overlapping (20–25 bp) PCR fragments amplified using Q5 High-Fidelity DNA Polymerase (NEB). .. Modified TALE constructs were created using PCR fragments amplified from pAAV-CW3SL-EGFP (Addgene #61463), a gift from Bong-Kiun Kaang , and constructs contained in the TALE Toolbox (Addgene kit #1000000019), a gift from Feng Zhang. ..

    Amplification:

    Article Title: Tet1 Isoforms Differentially Regulate Gene Expression, Synaptic Transmission, and Memory in the Mammalian Brain
    Article Snippet: Briefly, primers were designed using NEBuilder Assembly Tool (v1.12.18) to generate overlapping (20–25 bp) PCR fragments amplified using Q5 High-Fidelity DNA Polymerase (NEB). .. Modified TALE constructs were created using PCR fragments amplified from pAAV-CW3SL-EGFP (Addgene #61463), a gift from Bong-Kiun Kaang , and constructs contained in the TALE Toolbox (Addgene kit #1000000019), a gift from Feng Zhang. ..

    other:

    Article Title: SPARCLE, a p53-induced lncRNA, controls apoptosis after genotoxic stress by promoting PARP-1 cleavage.
    Article Snippet: Cells were transfected with 2 mg of each TALEN using Lipofectamine 2000 (11668019, ThermoFisher Scientific) and plated by limiting dilution in 100mmdishes 48 hr post-transfection.

    Article Title: Angulin-1 seals tricellular contacts independently of tricellulin and claudins
    Article Snippet: Stable cell lines were selected by treatment with 500 μg/ml G-418 (Nacalai Tesque).

    TALENs:

    Article Title: SPARCLE , a p53-induced lncRNA, controls apoptosis after genotoxic stress by promoting PARP-1 cleavage
    Article Snippet: .. TALENs were generated using the TALE Toolbox kit (1000000019, Addgene). .. Cells were transfected with 2 μg of each TALEN using Lipofectamine 2000 (11668019, ThermoFisher Scientific) and plated by limiting dilution in 100 mm dishes 48 hr post-transfection.

    Generated:

    Article Title: SPARCLE , a p53-induced lncRNA, controls apoptosis after genotoxic stress by promoting PARP-1 cleavage
    Article Snippet: .. TALENs were generated using the TALE Toolbox kit (1000000019, Addgene). .. Cells were transfected with 2 μg of each TALEN using Lipofectamine 2000 (11668019, ThermoFisher Scientific) and plated by limiting dilution in 100 mm dishes 48 hr post-transfection.



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    cell transcriptomic sequencing dataset  (10X Genomics)
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    10X Genomics cell transcriptomic sequencing dataset
    <t>Transcriptomic</t> and TME characteristics associated with ECMSig in TCGA-GBM cohort (A) Volcano plot showing DEGs between ECMSig-high and ECMSig-low groups. Red dots: upregulated in high-risk; blue dots: upregulated in low-risk. Benjamini-Hochberg adjusted. (B) Gene set enrichment analysis (GSEA) plots showing enrichment of hallmark pathways. Pathways enriched in ECMSig-high and ECMSig-low groups are shown with their running enrichment scores (ESs) and ranked gene lists. Benjamini-Hochberg adjusted. (C) Heatmap showing the activity scores of selected oncogenic and tumor-related signaling pathways (rows) across TCGA-GBM samples (columns), annotated by ECMSig group and ECMSig score. Red indicates high activity, blue indicates low activity. ∗ p < 0.05. Wilcoxon signed-rank test. (D) Heatmap depicting the estimated infiltration levels of various immune and stromal cell types (rows) in TCGA-GBM samples (columns), stratified by ECMSig group and score. Red indicates high infiltration, blue indicates low infiltration. Cells significantly highly infiltrated in ECMSig-high are labeled in red, and those high in ECMSig-low group are in blue. ∗q < 0.05, ∗∗q < 0.01, ∗∗∗q < 0.001. Wilcoxon signed-rank test. Benjamini-Hochberg adjusted. (E and F) Scatterplots showing the spearman correlation between ECMSig score and (E) Macrophage_XCELL infiltration score and (F) immune_score_XCELL. The blue line represents the linear regression fit with 95% confidence interval bands. Spearman correlation test.
    Cell Transcriptomic Sequencing Dataset, supplied by 10X Genomics, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/cell+transcriptomic+sequencing+dataset/cell+copy+genomic+number+profiling+single+technologies/pmc13197639-319-1-9
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    cell transcriptomic sequencing dataset - by Bioz Stars, 2026-11
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    Image Search Results


    Transcriptomic and TME characteristics associated with ECMSig in TCGA-GBM cohort (A) Volcano plot showing DEGs between ECMSig-high and ECMSig-low groups. Red dots: upregulated in high-risk; blue dots: upregulated in low-risk. Benjamini-Hochberg adjusted. (B) Gene set enrichment analysis (GSEA) plots showing enrichment of hallmark pathways. Pathways enriched in ECMSig-high and ECMSig-low groups are shown with their running enrichment scores (ESs) and ranked gene lists. Benjamini-Hochberg adjusted. (C) Heatmap showing the activity scores of selected oncogenic and tumor-related signaling pathways (rows) across TCGA-GBM samples (columns), annotated by ECMSig group and ECMSig score. Red indicates high activity, blue indicates low activity. ∗ p < 0.05. Wilcoxon signed-rank test. (D) Heatmap depicting the estimated infiltration levels of various immune and stromal cell types (rows) in TCGA-GBM samples (columns), stratified by ECMSig group and score. Red indicates high infiltration, blue indicates low infiltration. Cells significantly highly infiltrated in ECMSig-high are labeled in red, and those high in ECMSig-low group are in blue. ∗q < 0.05, ∗∗q < 0.01, ∗∗∗q < 0.001. Wilcoxon signed-rank test. Benjamini-Hochberg adjusted. (E and F) Scatterplots showing the spearman correlation between ECMSig score and (E) Macrophage_XCELL infiltration score and (F) immune_score_XCELL. The blue line represents the linear regression fit with 95% confidence interval bands. Spearman correlation test.

    Journal: iScience

    Article Title: Multi-omics profiling-derived signature links cellular ecosystem to glioblastoma prognosis

    doi: 10.1016/j.isci.2026.115982

    Figure Lengend Snippet: Transcriptomic and TME characteristics associated with ECMSig in TCGA-GBM cohort (A) Volcano plot showing DEGs between ECMSig-high and ECMSig-low groups. Red dots: upregulated in high-risk; blue dots: upregulated in low-risk. Benjamini-Hochberg adjusted. (B) Gene set enrichment analysis (GSEA) plots showing enrichment of hallmark pathways. Pathways enriched in ECMSig-high and ECMSig-low groups are shown with their running enrichment scores (ESs) and ranked gene lists. Benjamini-Hochberg adjusted. (C) Heatmap showing the activity scores of selected oncogenic and tumor-related signaling pathways (rows) across TCGA-GBM samples (columns), annotated by ECMSig group and ECMSig score. Red indicates high activity, blue indicates low activity. ∗ p < 0.05. Wilcoxon signed-rank test. (D) Heatmap depicting the estimated infiltration levels of various immune and stromal cell types (rows) in TCGA-GBM samples (columns), stratified by ECMSig group and score. Red indicates high infiltration, blue indicates low infiltration. Cells significantly highly infiltrated in ECMSig-high are labeled in red, and those high in ECMSig-low group are in blue. ∗q < 0.05, ∗∗q < 0.01, ∗∗∗q < 0.001. Wilcoxon signed-rank test. Benjamini-Hochberg adjusted. (E and F) Scatterplots showing the spearman correlation between ECMSig score and (E) Macrophage_XCELL infiltration score and (F) immune_score_XCELL. The blue line represents the linear regression fit with 95% confidence interval bands. Spearman correlation test.

    Article Snippet: The single-cell transcriptomic sequencing dataset utilizing technology from the 10X Genomics platform was available under the accession number GEO: GSE182109 at the Gene Expression Omnibus (GEO) repository.

    Techniques: Activity Assay, Protein-Protein interactions, Labeling

    Single-cell RNA sequencing analysis revealing ECMSig expression across cell types and identification of prognostically relevant cell states in GBM (A) UMAP visualization of major cell types identified in GBM scRNA-seq data. (B) Dot plot showing the scaled average expression (color intensity) and percentage of cells expressing (dot size) canonical marker genes for each major cell type. (C) Dot plot showing the scaled average expression and percentage of cells expressing the seven ECMSig genes across major cell types. (D) UMAP plots showing the expression levels of individual ECMSig genes and overall ECMSig score across all cells. (E–G) UMAP plots illustrating Scissor-identified prognostically unfavorable (Scissor_Pos, red dashed circle) and favorable (Scissor_Neg, blue dashed circle; Scissor_Others, gray) cell subpopulations within (E) tumor cells, (F) myeloid cells, and (G) endothelial cells. (H–K) Violin plots comparing ECMSig scores among tumor cells grouped by Scissor status (H) and tumor type (I), and myeloid cells (J) or endothelial cells (K) grouped by Scissor status. ∗∗∗∗ p < 0.0001. Wilcoxon signed-rank test. (L) Dot plot showing differentially expressed marker genes between myeloid Scissor_Pos and other myeloid cells. Dot size indicates the fraction of cells in the group expressing the gene; color indicates average expression level.

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    doi: 10.1016/j.isci.2026.115982

    Figure Lengend Snippet: Single-cell RNA sequencing analysis revealing ECMSig expression across cell types and identification of prognostically relevant cell states in GBM (A) UMAP visualization of major cell types identified in GBM scRNA-seq data. (B) Dot plot showing the scaled average expression (color intensity) and percentage of cells expressing (dot size) canonical marker genes for each major cell type. (C) Dot plot showing the scaled average expression and percentage of cells expressing the seven ECMSig genes across major cell types. (D) UMAP plots showing the expression levels of individual ECMSig genes and overall ECMSig score across all cells. (E–G) UMAP plots illustrating Scissor-identified prognostically unfavorable (Scissor_Pos, red dashed circle) and favorable (Scissor_Neg, blue dashed circle; Scissor_Others, gray) cell subpopulations within (E) tumor cells, (F) myeloid cells, and (G) endothelial cells. (H–K) Violin plots comparing ECMSig scores among tumor cells grouped by Scissor status (H) and tumor type (I), and myeloid cells (J) or endothelial cells (K) grouped by Scissor status. ∗∗∗∗ p < 0.0001. Wilcoxon signed-rank test. (L) Dot plot showing differentially expressed marker genes between myeloid Scissor_Pos and other myeloid cells. Dot size indicates the fraction of cells in the group expressing the gene; color indicates average expression level.

    Article Snippet: The single-cell transcriptomic sequencing dataset utilizing technology from the 10X Genomics platform was available under the accession number GEO: GSE182109 at the Gene Expression Omnibus (GEO) repository.

    Techniques: Single Cell, RNA Sequencing, Expressing, Marker

    Spatial transcriptomic analysis revealing co-localization of ECMSig, hypoxia, Scissor-Positive cells, and pericytes in GBM (A) Spatial feature plots for four GBM samples. Each row represents a sample. Columns show spatial heatmaps of: ECMSig score, hypoxia signature score, tumor Scissor_Pos signature score, myeloid Scissor_Pos signature score, endothelial Scissor Pos signature score, and pericyte marker signature score. Color scale indicates scaled expression or score (low to high). Each dot represents a spatial barcoded spot.

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    Article Title: Multi-omics profiling-derived signature links cellular ecosystem to glioblastoma prognosis

    doi: 10.1016/j.isci.2026.115982

    Figure Lengend Snippet: Spatial transcriptomic analysis revealing co-localization of ECMSig, hypoxia, Scissor-Positive cells, and pericytes in GBM (A) Spatial feature plots for four GBM samples. Each row represents a sample. Columns show spatial heatmaps of: ECMSig score, hypoxia signature score, tumor Scissor_Pos signature score, myeloid Scissor_Pos signature score, endothelial Scissor Pos signature score, and pericyte marker signature score. Color scale indicates scaled expression or score (low to high). Each dot represents a spatial barcoded spot.

    Article Snippet: The single-cell transcriptomic sequencing dataset utilizing technology from the 10X Genomics platform was available under the accession number GEO: GSE182109 at the Gene Expression Omnibus (GEO) repository.

    Techniques: Marker, Expressing