aav packaging plasmid  (TaKaRa)

Journal: bioRxiv

Article Title: Generation and characterization of a tamoxifen-inducible, Cre driver rat for transgene expression in microglia

doi: 10.1101/2025.07.17.665447

Figure Lengend Snippet: (A) Primary cortical neurons were used to test our CreERT2 (Addgene: #210415) and DIO-mCherry constructs delivered using AAV transductions. (B) AAV DIO-mCherry produces minimal Cre-independent mCherry expression in PCNs. (C) Co-delivery of AAV-iCre (Addgene: #210416) and AAV DIO-mCherry to PCNs induces robust mCherry expression in PCNs. (D) AAV-CreERT2 and AAV DIO-mCherry co-transduction induces minimal tamoxifen-independent mCherry expression in PCNs, while addition of 4-hydroxytamoxifen (1µM) following the same AAV-transduction generates robust mCherry expression. (Scale Bar: 200 µm).

Article Snippet: The coding region for CreERT2 was amplified from MSCV CreERT2 puro (a gift from Tyler Jacks; Addgene; #22776) and inserted into an AAV packaging plasmid flanked by the EF1α promoter and the poly-adenylation signal from bovine growth hormone using ligation-independent cloning (In-Fusion, Clontech).

Techniques: Construct, Expressing, Transduction

Journal: bioRxiv

Article Title: Generation and characterization of a tamoxifen-inducible, Cre driver rat for transgene expression in microglia

doi: 10.1101/2025.07.17.665447

Figure Lengend Snippet: (A) The DIO-mCherry rat uses human EF1α promoter to drive expression of mCherry fluorescent protein along with the woodchuck post transcriptional regulatory element (WPRE) and the human growth hormone polyadenylation signal (hGHpA). (A) The primer positions for a PCR genotyping assay spanning the 5’ junction of the mCherry cassette is noted by the black arrows and the positions of the primers (blue) and probe (green arrow) for a qPCR-based genotyping assay are also noted. ( B ) Copy number for WT, hemizygous (Hem), and homozygous (Hom) animals. The ratio is transgene copy number to the genomic gene (Ggt1) in genomic DNA samples isolated from the pups of two separate Hem × Hem crosses. The individual ratios of templates (DIO-mCherry/Ggt1) fall into three quanta (0, 0.5, and 1.0). (C) Amplicons (640 bp) produced by the 5’ junction PCR identify wild type (no bands) or carriers hemizygous and homozygous. (D) Images from the striatum of hemizygous DIO-mCherry rats injected with an AAV virus expressing Cre recombinase after two weeks. Low magnification (top row; scale bar: 500 μm) and higher magnification (bottom row; scale bar: 50 μm). Cre immunoreactivity (white) and mCherry epifluorescence (red) colocalize in both sets of images. Overlap Coefficient (OC)=0.90.

Article Snippet: The coding region for CreERT2 was amplified from MSCV CreERT2 puro (a gift from Tyler Jacks; Addgene; #22776) and inserted into an AAV packaging plasmid flanked by the EF1α promoter and the poly-adenylation signal from bovine growth hormone using ligation-independent cloning (In-Fusion, Clontech).

Techniques: Expressing, Genotyping Assay, Isolation, Produced, Injection, Virus

Journal: Frontiers in Immunology

Article Title: Long-term protective potency of AAV vector-based SARS-CoV-2 prophylaxis in mice and non-human primates

doi: 10.3389/fimmu.2026.1829230

Figure Lengend Snippet: Analysis of serum P2C5-Fc concentration, neutralizing activity, and immunogenicity in mice following i.m. administration of rAAV-P2C5-Fc. (A) Study design. (B) Serum levels of the P2C5-Fc antibody following i.m. administration of rAAV-P2C5-Fc at a dose of a 1.0 × 10 13 gc/kg in mice. (C) NAb activity titers against SARS-CoV-2. (D) Antibody titers against AAV-DJ capsid proteins and the expressed P2C5-Fc antibody.

Article Snippet: The rAAV-P2C5-Fc vector construct was produced using a three-plasmid AAV-DJ packaging system (Cell Biolabs, USA) by transient transfection of HEK293 cells (obtained from the cell collection of the N.F.

Techniques: Concentration Assay, Activity Assay, Immunopeptidomics