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px330 spcas9 grna vector  (Addgene inc)


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    Structured Review

    Addgene inc px330 spcas9 grna vector
    Px330 Spcas9 Grna Vector, supplied by Addgene inc, used in various techniques. Bioz Stars score: 96/100, based on 2997 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/px330+grna+vector/pX330-U6-Chimeric_BB-CBh-hSpCas9+(Plasmid+%2342230)/bio_rxiv__2025__05__14__654043-182-37-40
    Average 96 stars, based on 2997 article reviews
    px330 spcas9 grna vector - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Synthesized:

    Article Title: Spatial organizations of heterochromatin underpin nuclear structural integrity of ventricular cardiomyocytes against mechanical stress.
    Article Snippet: In brief Cardiomyocytes exhibit characteristic prominent spatial organizations of heterochromatin (SOH).. Fujiwara et al. found that the replacement of endogenous H2B with H2B-mCherry disrupts SOH, leading to extreme elongation of ventricular cardiomyocyte nuclei, resulting in severe heart failure, demonstrating that SOH play a structural role in maintaining nuclear shape against mechanical stress.

    Clone Assay:

    Article Title: Spatial organizations of heterochromatin underpin nuclear structural integrity of ventricular cardiomyocytes against mechanical stress.
    Article Snippet: In brief Cardiomyocytes exhibit characteristic prominent spatial organizations of heterochromatin (SOH).. Fujiwara et al. found that the replacement of endogenous H2B with H2B-mCherry disrupts SOH, leading to extreme elongation of ventricular cardiomyocyte nuclei, resulting in severe heart failure, demonstrating that SOH play a structural role in maintaining nuclear shape against mechanical stress.

    Plasmid Preparation:

    Article Title: Spatial organizations of heterochromatin underpin nuclear structural integrity of ventricular cardiomyocytes against mechanical stress.
    Article Snippet: In brief Cardiomyocytes exhibit characteristic prominent spatial organizations of heterochromatin (SOH).. Fujiwara et al. found that the replacement of endogenous H2B with H2B-mCherry disrupts SOH, leading to extreme elongation of ventricular cardiomyocyte nuclei, resulting in severe heart failure, demonstrating that SOH play a structural role in maintaining nuclear shape against mechanical stress.

    Article Title: Mechanistic basis of gene regulation by SRCAP and H2A.Z
    Article Snippet: .. A SRCAP gRNA designed using CRISPOR to target the C-terminal part of mouse SRCAP was added in pX330-gRNA vector (pX330-gRNA was a gift from Charles P. Lai (Addgene plasmid #158973)) by digestion with BbsI to build pX330_SRCAP. .. The Ypet_AID_P2A_Hygromycin-resistance cassette was amplified with primers containing 60bp homology arms corresponding to the carboxy-terminal part of SRCAP and 2μg of amplicons were co-transfected in 1.5×10 6 CGR8 ESC together with 2 μg of pX330-SRCAP using Lipofectamine 3000 (ThermoFisher Scientific #L3000008).

    Selection:

    Article Title: Spatial organizations of heterochromatin underpin nuclear structural integrity of ventricular cardiomyocytes against mechanical stress.
    Article Snippet: In brief Cardiomyocytes exhibit characteristic prominent spatial organizations of heterochromatin (SOH).. Fujiwara et al. found that the replacement of endogenous H2B with H2B-mCherry disrupts SOH, leading to extreme elongation of ventricular cardiomyocyte nuclei, resulting in severe heart failure, demonstrating that SOH play a structural role in maintaining nuclear shape against mechanical stress.

    Cell Culture:

    Article Title: Spatial organizations of heterochromatin underpin nuclear structural integrity of ventricular cardiomyocytes against mechanical stress.
    Article Snippet: In brief Cardiomyocytes exhibit characteristic prominent spatial organizations of heterochromatin (SOH).. Fujiwara et al. found that the replacement of endogenous H2B with H2B-mCherry disrupts SOH, leading to extreme elongation of ventricular cardiomyocyte nuclei, resulting in severe heart failure, demonstrating that SOH play a structural role in maintaining nuclear shape against mechanical stress.



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    Image Search Results


    (A) Domain structure of the two ADAR1 isoforms, p150 and p110. (B) Schematic representation of the CRISPR/Cas9-mediated disruption of ADAR1 p150 and p150/p110. The target sequence and corresponding vectors containing either a reverse-oriented puromycin- or neomycin- resistance cassette are illustrated. (C) Western blot analysis of ADAR1 isoforms. Whole-cell extracts from WT, p150 KO clones (#1, #2, #3), and p150/p110 KO clones (#1, #2, #3) were resolved by 10% SDS-PAGE. GAPDH was used as a loading control. (D) Clustering analysis of genes exhibiting A- to-I editing. Based on inosine peak scores (fold-enrichment ratio) in WT and p150 KO cells, genes were categorized as strongly p150 dependent (WT/(p150KO+1)≧4; purple), mildly p150 dependent (1<WT/(p150KO+1)<4; orange), p150 independent (WT/(p150KO+1)<1; pink), and newly detected upon p150 loss, i.e., p150KO-specific (WT=0, p150KO>1, cyan). Asterisk (*) indicates the genes categorized as p150/p110 KO-specific (WT=0, p150KO=0, p150/p110KO>1).

    Journal: bioRxiv

    Article Title: Epitranscriptome-wide profiling identifies RNA editing events regulated by ADAR1 that are associated with DNA repair mechanisms in human TK6 cells

    doi: 10.1101/2025.07.11.664482

    Figure Lengend Snippet: (A) Domain structure of the two ADAR1 isoforms, p150 and p110. (B) Schematic representation of the CRISPR/Cas9-mediated disruption of ADAR1 p150 and p150/p110. The target sequence and corresponding vectors containing either a reverse-oriented puromycin- or neomycin- resistance cassette are illustrated. (C) Western blot analysis of ADAR1 isoforms. Whole-cell extracts from WT, p150 KO clones (#1, #2, #3), and p150/p110 KO clones (#1, #2, #3) were resolved by 10% SDS-PAGE. GAPDH was used as a loading control. (D) Clustering analysis of genes exhibiting A- to-I editing. Based on inosine peak scores (fold-enrichment ratio) in WT and p150 KO cells, genes were categorized as strongly p150 dependent (WT/(p150KO+1)≧4; purple), mildly p150 dependent (11, cyan). Asterisk (*) indicates the genes categorized as p150/p110 KO-specific (WT=0, p150KO=0, p150/p110KO>1).

    Article Snippet: The vectors pX330-gRNA/p150 (6 μg) and the p150 target plasmid (2 μg) were transfected into TK6 cells by a NEPA21 electroporator (Nepa Gene Co. Ltd.) following the manufacturer’s instructions.

    Techniques: CRISPR, Disruption, Sequencing, Western Blot, Clone Assay, SDS Page, Control

    Read coverage profiles are shown for representative DNA repair-associated genes: (A) ATM (3′UTR), (B) POLH (3′UTR), (C) POLH (intron between exon4-5), (D) ATR (intron between exon1-2), (E) FANCA (intron between exon5-6), (F), FANCA (intron between exon14-15), and (G) XPA (intron between exon5-6) in wild-type (cyan), p150 KO (pink), and p150/p110 KO cells (green). The y-axis represents read counts and the x-axis indicates genomic coordinates. The scale bar corresponds to 1 kb. “Coverage data range” in the top-right corner of each panel indicates the maximum coverage value. Colored regions within coverage tracks highlight A-to-I RNA editing sites. Strand-specific colors compositions are shown: green (Adenosine) and orange (Guanosine) for forward reads; red (Thymidine) and blue (Cytidine) for reverse reads, facilitating visual estimation of nucleotide ratios at individual position.

    Journal: bioRxiv

    Article Title: Epitranscriptome-wide profiling identifies RNA editing events regulated by ADAR1 that are associated with DNA repair mechanisms in human TK6 cells

    doi: 10.1101/2025.07.11.664482

    Figure Lengend Snippet: Read coverage profiles are shown for representative DNA repair-associated genes: (A) ATM (3′UTR), (B) POLH (3′UTR), (C) POLH (intron between exon4-5), (D) ATR (intron between exon1-2), (E) FANCA (intron between exon5-6), (F), FANCA (intron between exon14-15), and (G) XPA (intron between exon5-6) in wild-type (cyan), p150 KO (pink), and p150/p110 KO cells (green). The y-axis represents read counts and the x-axis indicates genomic coordinates. The scale bar corresponds to 1 kb. “Coverage data range” in the top-right corner of each panel indicates the maximum coverage value. Colored regions within coverage tracks highlight A-to-I RNA editing sites. Strand-specific colors compositions are shown: green (Adenosine) and orange (Guanosine) for forward reads; red (Thymidine) and blue (Cytidine) for reverse reads, facilitating visual estimation of nucleotide ratios at individual position.

    Article Snippet: The vectors pX330-gRNA/p150 (6 μg) and the p150 target plasmid (2 μg) were transfected into TK6 cells by a NEPA21 electroporator (Nepa Gene Co. Ltd.) following the manufacturer’s instructions.

    Techniques:

    (A) Gene expression levels of ATM, ATR , and FANCA were quantified by RT-qPCR in WT (black), p150 KO (gray), and p150/p110 KO (white) cells. Data represent six independent experiments (n = 6). Statistical significance was determined by Student’s t-test (p < 0.05). (B) RNA-seq read coverage profiles of the XPA gene in WT#1, WT#2, p150 KO#1, p150 KO#2, p150/p110 KO#1, and p150/p110 KO#2 cells. Arrows indicate novel splicing peaks between exon 5 and 6 of the main splicing variant. (C) Detection of XPA splicing variants. PCR was performed using cDNA from WT#1, p150 KO#1, p150 KO#2, p150/p110 KO#1, and p150/p110 KO#2, with primers targeting exons 2–6 and 5–6. Amplified products were resolved on 1.0% agarose gels. For amplification between exons 2–6, expected band sizes were 946 bp for variant 1 and 1,260 bp for a potential alternative variant. For amplification between exons 5–6, expected sizes were 544 bp for variant 1 and 964 bp for alternative variants.

    Journal: bioRxiv

    Article Title: Epitranscriptome-wide profiling identifies RNA editing events regulated by ADAR1 that are associated with DNA repair mechanisms in human TK6 cells

    doi: 10.1101/2025.07.11.664482

    Figure Lengend Snippet: (A) Gene expression levels of ATM, ATR , and FANCA were quantified by RT-qPCR in WT (black), p150 KO (gray), and p150/p110 KO (white) cells. Data represent six independent experiments (n = 6). Statistical significance was determined by Student’s t-test (p < 0.05). (B) RNA-seq read coverage profiles of the XPA gene in WT#1, WT#2, p150 KO#1, p150 KO#2, p150/p110 KO#1, and p150/p110 KO#2 cells. Arrows indicate novel splicing peaks between exon 5 and 6 of the main splicing variant. (C) Detection of XPA splicing variants. PCR was performed using cDNA from WT#1, p150 KO#1, p150 KO#2, p150/p110 KO#1, and p150/p110 KO#2, with primers targeting exons 2–6 and 5–6. Amplified products were resolved on 1.0% agarose gels. For amplification between exons 2–6, expected band sizes were 946 bp for variant 1 and 1,260 bp for a potential alternative variant. For amplification between exons 5–6, expected sizes were 544 bp for variant 1 and 964 bp for alternative variants.

    Article Snippet: The vectors pX330-gRNA/p150 (6 μg) and the p150 target plasmid (2 μg) were transfected into TK6 cells by a NEPA21 electroporator (Nepa Gene Co. Ltd.) following the manufacturer’s instructions.

    Techniques: Gene Expression, Quantitative RT-PCR, RNA Sequencing, Variant Assay, Amplification