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specificity cas9  (Addgene inc)


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    Structured Review

    Addgene inc specificity cas9
    Specificity Cas9, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/specificity+cas9/pCAG-eCas9-GFP-U6-gRNA+(Plasmid+%2379145)/pm41422089-164-9-12
    Average 94 stars, based on 55 article reviews
    specificity cas9 - by Bioz Stars, 2026-10
    94/100 stars

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    Related Articles

    Plasmid Preparation:

    Article Title: The Ku80-p53-SIRT1 axis in DNA damage response contributes to sporadic and familial ALS and FTD.
    Article Snippet: .. A plasmid expressing both the sgRNA (5’-GGTAAGCTTCGAGCAGCTGATGG-3’) and enhanced specificity Cas9 (pCAG-eCas9-GFP-U6-gRNA, Addgene #79145) was used. .. Two million iPSCs were nucleofected with 1 μg of the CRISPR-Cas9 plasmid and 4 μl of the ssODN using a 4D-Nucleofector (Amaxa, program CA137).

    Article Title: The Ku80-p53-SIRT1 axis in DNA damage response contributes to sporadic and familial ALS and FTD
    Article Snippet: .. A plasmid expressing both the sgRNA (5’-GGTAAGCTTCGAGCAGCTGATGG-3’) and enhanced specificity Cas9 (pCAG-eCas9-GFP-U6-gRNA, Addgene #79145) was used. .. Two million iPSCs were nucleofected with 1 μg of the CRISPR-Cas9 plasmid and 4 μl of the ssODN using a 4D-Nucleofector (Amaxa, program CA137).

    Article Title: Nonsense, but not frameshift, truncating mutations result in Cyclin D2 stabilisation in an induced pluripotent stem cell model of MPPH
    Article Snippet: Cultures were regularly tested for pluripotency by Immunofluoresence of Oct3/4 and SSEA, and mycoplasma using MycoStrips (Invitrogen). .. A single-guide RNA targeting the final exon of CCND2 (5’-ACGTGACGGATCCAAGTCGG-3’) was ligated in to either the GFP-tagged CRISPR-Cas9 px458 plasmid (Addgene #48138) or the enhanced specificity Cas9 (eSpCas9, Addgene #71814) following digestion with BbsI restriction enzyme (Thermo Fisher Scientific). ..

    Expressing:

    Article Title: The Ku80-p53-SIRT1 axis in DNA damage response contributes to sporadic and familial ALS and FTD.
    Article Snippet: .. A plasmid expressing both the sgRNA (5’-GGTAAGCTTCGAGCAGCTGATGG-3’) and enhanced specificity Cas9 (pCAG-eCas9-GFP-U6-gRNA, Addgene #79145) was used. .. Two million iPSCs were nucleofected with 1 μg of the CRISPR-Cas9 plasmid and 4 μl of the ssODN using a 4D-Nucleofector (Amaxa, program CA137).

    Article Title: The Ku80-p53-SIRT1 axis in DNA damage response contributes to sporadic and familial ALS and FTD
    Article Snippet: .. A plasmid expressing both the sgRNA (5’-GGTAAGCTTCGAGCAGCTGATGG-3’) and enhanced specificity Cas9 (pCAG-eCas9-GFP-U6-gRNA, Addgene #79145) was used. .. Two million iPSCs were nucleofected with 1 μg of the CRISPR-Cas9 plasmid and 4 μl of the ssODN using a 4D-Nucleofector (Amaxa, program CA137).

    CRISPR:

    Article Title: Nonsense, but not frameshift, truncating mutations result in Cyclin D2 stabilisation in an induced pluripotent stem cell model of MPPH
    Article Snippet: Cultures were regularly tested for pluripotency by Immunofluoresence of Oct3/4 and SSEA, and mycoplasma using MycoStrips (Invitrogen). .. A single-guide RNA targeting the final exon of CCND2 (5’-ACGTGACGGATCCAAGTCGG-3’) was ligated in to either the GFP-tagged CRISPR-Cas9 px458 plasmid (Addgene #48138) or the enhanced specificity Cas9 (eSpCas9, Addgene #71814) following digestion with BbsI restriction enzyme (Thermo Fisher Scientific). ..



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    Membrane anchor mutants of Miro1/2 rescue Miro DKO. (A) Scheme of Miro1 and <t>Miro2</t> residues that were exchanged with the corresponding residues of VAMP1B and Bcl-xL, respectively. Residues highlighted in green and blue are part of the transmembrane regions and those in red of the intermembrane space (IMS). Residues in light blue color are derived from VAMP1B and face the cytosol. Dm: Drosophila melanogaster , Hs: Homo sapiens . (B) Western blot analysis of protein levels of Miro, Halo–Miro, Myo19 and β-actin in WT, Miro DKO and Miro DKO cells expressing different Miro constructs as indicated. (C) Quantification of Myo19 protein levels in the different cell lines as indicated (normalized to β-actin levels). Results are mean±s.e.m. ( n =3). * P <0.05; ** P <0.01; *** P <0.001 (two-sample unpaired t -test). (D,E) Confocal images of Miro DKO cells rescued with Halo-tagged Miro constructs (D) and Halo-tagged mutant Miro constructs (E) that were transfected with GFP–Myo19. Mitochondria were stained with Mitotracker Orange. n =20–30 cells per experiment ( n =3 experiments). Scale bars, 10 µm. (F) Miro DKO cells rescued with either Halo–Miro1, Halo–Miro2, Halo–Miro1mutant or Halo–Miro2mutant were lysed. The lysates were subjected to Halo–Trap agarose pull-downs. Western blot analysis of inputs and eluates for Miro, Myo19, Mic60, Sam50, Mtx3 and GAPDH are shown. Images in F are representative of four repeats.
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    Image Search Results


    Membrane anchor mutants of Miro1/2 rescue Miro DKO. (A) Scheme of Miro1 and Miro2 residues that were exchanged with the corresponding residues of VAMP1B and Bcl-xL, respectively. Residues highlighted in green and blue are part of the transmembrane regions and those in red of the intermembrane space (IMS). Residues in light blue color are derived from VAMP1B and face the cytosol. Dm: Drosophila melanogaster , Hs: Homo sapiens . (B) Western blot analysis of protein levels of Miro, Halo–Miro, Myo19 and β-actin in WT, Miro DKO and Miro DKO cells expressing different Miro constructs as indicated. (C) Quantification of Myo19 protein levels in the different cell lines as indicated (normalized to β-actin levels). Results are mean±s.e.m. ( n =3). * P <0.05; ** P <0.01; *** P <0.001 (two-sample unpaired t -test). (D,E) Confocal images of Miro DKO cells rescued with Halo-tagged Miro constructs (D) and Halo-tagged mutant Miro constructs (E) that were transfected with GFP–Myo19. Mitochondria were stained with Mitotracker Orange. n =20–30 cells per experiment ( n =3 experiments). Scale bars, 10 µm. (F) Miro DKO cells rescued with either Halo–Miro1, Halo–Miro2, Halo–Miro1mutant or Halo–Miro2mutant were lysed. The lysates were subjected to Halo–Trap agarose pull-downs. Western blot analysis of inputs and eluates for Miro, Myo19, Mic60, Sam50, Mtx3 and GAPDH are shown. Images in F are representative of four repeats.

    Journal: Journal of Cell Science

    Article Title: Regulation of mitochondrial cristae organization by Myo19, Miro1 and Miro2, and metaxin 3

    doi: 10.1242/jcs.263637

    Figure Lengend Snippet: Membrane anchor mutants of Miro1/2 rescue Miro DKO. (A) Scheme of Miro1 and Miro2 residues that were exchanged with the corresponding residues of VAMP1B and Bcl-xL, respectively. Residues highlighted in green and blue are part of the transmembrane regions and those in red of the intermembrane space (IMS). Residues in light blue color are derived from VAMP1B and face the cytosol. Dm: Drosophila melanogaster , Hs: Homo sapiens . (B) Western blot analysis of protein levels of Miro, Halo–Miro, Myo19 and β-actin in WT, Miro DKO and Miro DKO cells expressing different Miro constructs as indicated. (C) Quantification of Myo19 protein levels in the different cell lines as indicated (normalized to β-actin levels). Results are mean±s.e.m. ( n =3). * P <0.05; ** P <0.01; *** P <0.001 (two-sample unpaired t -test). (D,E) Confocal images of Miro DKO cells rescued with Halo-tagged Miro constructs (D) and Halo-tagged mutant Miro constructs (E) that were transfected with GFP–Myo19. Mitochondria were stained with Mitotracker Orange. n =20–30 cells per experiment ( n =3 experiments). Scale bars, 10 µm. (F) Miro DKO cells rescued with either Halo–Miro1, Halo–Miro2, Halo–Miro1mutant or Halo–Miro2mutant were lysed. The lysates were subjected to Halo–Trap agarose pull-downs. Western blot analysis of inputs and eluates for Miro, Myo19, Mic60, Sam50, Mtx3 and GAPDH are shown. Images in F are representative of four repeats.

    Article Snippet: A human Miro2 target specific CRISPR-Cas9 knockout plasmid was purchased from Santa Cruz Biotechnology (sc-406496).

    Techniques: Membrane, Derivative Assay, Western Blot, Expressing, Construct, Mutagenesis, Transfection, Staining

    Ultrastructural analysis of mitochondria in various Miro DKO rescue cells. Representative TEM images are shown of mitochondria in HEK WT (A), Miro DKO (B), Miro DKO with Halo–Miro1 (MiroDKO-HaloMiro1) (C), Miro DKO with Halo–Miro2 (MiroDKO-HaloMiro2) (D), Miro DKO with Halo–Miro1 mutant (MiroDKO-HaloMiro1 mutant) (E) and Miro DKO with Halo–Miro2 mutant (MiroDKO-HaloMiro2 mutant) cells (F). In G, characteristic examples are shown of mitochondria with normal, intermediate and aberrant cristae organization. (H) Percentages of mitochondria with normal, intermediate and aberrant cristae organization in the indicated cell lines. n =200–250 mitochondria for WT and Miro DKO, n =150–175 mitochondria for the rescue Miro DKO cell lines. Scale bars: 500 nm.

    Journal: Journal of Cell Science

    Article Title: Regulation of mitochondrial cristae organization by Myo19, Miro1 and Miro2, and metaxin 3

    doi: 10.1242/jcs.263637

    Figure Lengend Snippet: Ultrastructural analysis of mitochondria in various Miro DKO rescue cells. Representative TEM images are shown of mitochondria in HEK WT (A), Miro DKO (B), Miro DKO with Halo–Miro1 (MiroDKO-HaloMiro1) (C), Miro DKO with Halo–Miro2 (MiroDKO-HaloMiro2) (D), Miro DKO with Halo–Miro1 mutant (MiroDKO-HaloMiro1 mutant) (E) and Miro DKO with Halo–Miro2 mutant (MiroDKO-HaloMiro2 mutant) cells (F). In G, characteristic examples are shown of mitochondria with normal, intermediate and aberrant cristae organization. (H) Percentages of mitochondria with normal, intermediate and aberrant cristae organization in the indicated cell lines. n =200–250 mitochondria for WT and Miro DKO, n =150–175 mitochondria for the rescue Miro DKO cell lines. Scale bars: 500 nm.

    Article Snippet: A human Miro2 target specific CRISPR-Cas9 knockout plasmid was purchased from Santa Cruz Biotechnology (sc-406496).

    Techniques: Mutagenesis