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adeno associated virus backbone  (Addgene inc)


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    Addgene inc adeno associated virus backbone
    Adeno Associated Virus Backbone, supplied by Addgene inc, used in various techniques. Bioz Stars score: 94/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/adeno associated virus backbone/product/Addgene inc
    Average 94 stars, based on 62 article reviews
    adeno associated virus backbone - by Bioz Stars, 2026-05
    94/100 stars

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    FIG. 2. CRISPR-CLONInG: Replacement of partial cargo sequence (Cre-comp) with the desired donor sequence on adeno-associated virus <t>(AAV)</t> vector (#60229; Addgene). (A) Schematic illustration of <t>AAV</t> <t>vector</t> with CRISPR cut sites (red scissors) at two ends of the Cre-comp segment. Guides (AAV-A and AAV-B) with high on-target scores were selected. (B) Cre-comp was cut out with ctRNP (Cas9-ctRNA) complex; donor for gene replacement (containing 15 bp AA replacement sequence, noted as ‘‘R,’’ sandwiched by HA) flanked with complementary Gibson overhangs of the adjacent AAV backbone was PCR-amplified from custom gene synthesized plasmid. (C) Assembled AAV-v1: donor template cloned into the customized AAV backbone via Gibson (HiFi) Assembly. (D) Excised AAV vector backbone (*3.63 kb) and Cre-comp (*2.73 kb; left); PCR amplified donor template (*0.8 kb) with Gibson overhangs (right). (E) After CRISPR-CLONInG, 15/16 clones showed correct vector assembly, confirmed by BbsI RE(s) diagnosis (two DNA fragments; black arrow); three clones further validated by Sanger sequencing. Resolved on 0.9% agarose gel.
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    FIG. 2. CRISPR-CLONInG: Replacement of partial cargo sequence (Cre-comp) with the desired donor sequence on adeno-associated virus (AAV) vector (#60229; Addgene). (A) Schematic illustration of AAV vector with CRISPR cut sites (red scissors) at two ends of the Cre-comp segment. Guides (AAV-A and AAV-B) with high on-target scores were selected. (B) Cre-comp was cut out with ctRNP (Cas9-ctRNA) complex; donor for gene replacement (containing 15 bp AA replacement sequence, noted as ‘‘R,’’ sandwiched by HA) flanked with complementary Gibson overhangs of the adjacent AAV backbone was PCR-amplified from custom gene synthesized plasmid. (C) Assembled AAV-v1: donor template cloned into the customized AAV backbone via Gibson (HiFi) Assembly. (D) Excised AAV vector backbone (*3.63 kb) and Cre-comp (*2.73 kb; left); PCR amplified donor template (*0.8 kb) with Gibson overhangs (right). (E) After CRISPR-CLONInG, 15/16 clones showed correct vector assembly, confirmed by BbsI RE(s) diagnosis (two DNA fragments; black arrow); three clones further validated by Sanger sequencing. Resolved on 0.9% agarose gel.

    Journal: The CRISPR journal

    Article Title: New Additions to the CRISPR Toolbox: CRISPR- CLONInG and CRISPR- CLIP for Donor Construction in Genome Editing.

    doi: 10.1089/crispr.2019.0062

    Figure Lengend Snippet: FIG. 2. CRISPR-CLONInG: Replacement of partial cargo sequence (Cre-comp) with the desired donor sequence on adeno-associated virus (AAV) vector (#60229; Addgene). (A) Schematic illustration of AAV vector with CRISPR cut sites (red scissors) at two ends of the Cre-comp segment. Guides (AAV-A and AAV-B) with high on-target scores were selected. (B) Cre-comp was cut out with ctRNP (Cas9-ctRNA) complex; donor for gene replacement (containing 15 bp AA replacement sequence, noted as ‘‘R,’’ sandwiched by HA) flanked with complementary Gibson overhangs of the adjacent AAV backbone was PCR-amplified from custom gene synthesized plasmid. (C) Assembled AAV-v1: donor template cloned into the customized AAV backbone via Gibson (HiFi) Assembly. (D) Excised AAV vector backbone (*3.63 kb) and Cre-comp (*2.73 kb; left); PCR amplified donor template (*0.8 kb) with Gibson overhangs (right). (E) After CRISPR-CLONInG, 15/16 clones showed correct vector assembly, confirmed by BbsI RE(s) diagnosis (two DNA fragments; black arrow); three clones further validated by Sanger sequencing. Resolved on 0.9% agarose gel.

    Article Snippet: CRISPR-CLONInG: Replacement of partial cargo sequence (Cre-comp) with the desired donor sequence on adeno-associated virus (AAV) vector (#60229; Addgene). (A) Schematic illustration of AAV vector with CRISPR cut sites (red scissors) at two ends of the Cre-comp segment.

    Techniques: CRISPR, Cloning, Sequencing, Virus, Plasmid Preparation, Synthesized, Clone Assay, Biomarker Discovery, Agarose Gel Electrophoresis