aav vector aav itr u6 sgrna backbone hsyn egfp kash wpre shortpa itr  (Addgene inc)

Journal: Biomolecules

Article Title: Transduction and Genome Editing of the Heart with Adeno-Associated Viral Vectors Loaded onto Electrospun Polydioxanone Nonwoven Fabrics.

doi: 10.3390/biom14040506

Figure Lengend Snippet: Figure 3. Release profile of AAV vectors from PDO electrospun nonwoven fabrics. Cumulative release of the vectors from the fabrics obtained by soaking in a solution containing 2.0 × 107 GCs of scAAV-GFP vectors. Data represent means ± SDs.

Article Snippet: Plasmid The gRNA cloning plasmid (gRNA_cloning vector), GFP-expression self-comple- mentary AAV plasmid (pscAAV-CAG-GFP), GFP-containing plasmid (pCAG-1BPNLSCas9-1BPNLS-2AGFP), Cas9-expression AAV plasmid (pAAV-nEFCas9), and AAV donor backbone plasmid (pAAV-rMERTK-HITI) were obtained from Addgene (Addgene 41824, 83279, 87109, 87115, and 87119, respectively).

Techniques:

Journal: Biomolecules

Article Title: Transduction and Genome Editing of the Heart with Adeno-Associated Viral Vectors Loaded onto Electrospun Polydioxanone Nonwoven Fabrics.

doi: 10.3390/biom14040506

Figure Lengend Snippet: Figure 5. Transduction to 10T1/2 cells, HepG2 cells, Hepa1-6 cells, and C2C12 cells on PDO elec- trospun nonwoven fabrics with AAV vectors. Fluorescence images of the transduced cells on the nonwoven fabrics. GFP expression rates are calculated from the number of GFP-positive cells. Data represent means ± SDs.

Article Snippet: Plasmid The gRNA cloning plasmid (gRNA_cloning vector), GFP-expression self-comple- mentary AAV plasmid (pscAAV-CAG-GFP), GFP-containing plasmid (pCAG-1BPNLSCas9-1BPNLS-2AGFP), Cas9-expression AAV plasmid (pAAV-nEFCas9), and AAV donor backbone plasmid (pAAV-rMERTK-HITI) were obtained from Addgene (Addgene 41824, 83279, 87109, 87115, and 87119, respectively).

Techniques: Transduction, Fluorescence, Expressing

Journal: Biomolecules

Article Title: Transduction and Genome Editing of the Heart with Adeno-Associated Viral Vectors Loaded onto Electrospun Polydioxanone Nonwoven Fabrics.

doi: 10.3390/biom14040506

Figure Lengend Snippet: Figure 7. Genome editing in mouse heart with attached electrospun PDO nonwoven fabrics with AAV vectors for gene knock-in through HITI. Schematic representation of (a) the Cas9/gRNA target site at exon 39 of the Myh6 locus and (b) genome-editing assays for GFP knock-in through HITI. The light-blue pentagon represents the Cas9/gRNA target sequence. The black line within the light-blue pentagon represents the Cas9 cleavage site. (c) A representative image of immunostaining of GFP- positive cells (green) with nucleic staining by Hoechst 33342 (blue), 14 days after the attachment of the fabrics to the heart below the fabrics with the AAV vectors for gene knock-in through HITI. The scale bar represents 200 µm.

Article Snippet: Plasmid The gRNA cloning plasmid (gRNA_cloning vector), GFP-expression self-comple- mentary AAV plasmid (pscAAV-CAG-GFP), GFP-containing plasmid (pCAG-1BPNLSCas9-1BPNLS-2AGFP), Cas9-expression AAV plasmid (pAAV-nEFCas9), and AAV donor backbone plasmid (pAAV-rMERTK-HITI) were obtained from Addgene (Addgene 41824, 83279, 87109, 87115, and 87119, respectively).

Techniques: Gene Knock-In, Knock-In, Sequencing, Immunostaining, Staining