Penn aav hsyn cre wpre hgh

Manufactured by Addgene

PENN-AAV-hSyn-Cre-WPRE-hGH is an adeno-associated virus (AAV) vector containing the Cre recombinase gene under the control of the human synapsin (hSyn) promoter, along with the Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) and the human growth hormone (hGH) polyadenylation signal. This vector is designed for Cre-mediated recombination in neuronal cells.

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Lab products found in correlation

Paav hsyn flex voltron st, by Addgene (2 mentions) Paav hsyn hchr2 h134r eyfp, by Addgene (1 mentions) P3000mp, by Thermo Fisher Scientific (1 mentions) D2650, by Merck Group (1 mentions) Aav hsyn dio hm3d gq mcherry, by Addgene (1 mentions) Aav hsyn egfp, by Addgene (1 mentions)

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AAV-hSyn-Cre-WPRE-hGH (2 citations)

3 protocols using penn aav hsyn cre wpre hgh

Optogenetic Neuronal Manipulation Toolkit

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pAAV-hsyn-flex-Voltron-ST (AAV1); Addgene, catalog no. 119036-AAV1; (at titer ≥2 × 10^–12 viral genomes ml^–1)
pAAV-hSyn-hChR2(H134R)-EYFP (AAV5); Addgene, catalog no. 26973-AAV5; (at titer ≥7 × 10^–12 viral genomes ml^–1)
pENN-AAV-hSyn-Cre-WPRE-hGH (AAV1); Addgene, catalog no. 05553-AAV1; (at titer ≥1 × 10^–13 viral genomes ml^–1)

Csillag V., Bizzozzero M.H., Noble J.C., Reinius B, & Fuzik J. (2023). Voltage-Seq: all-optical postsynaptic connectome-guided single-cell transcriptomics. Nature Methods, 20(9), 1409-1416.

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Viral Transduction and Calcium Imaging in Mouse Brain

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Male and female wild‐type C57BL/6Crl mice aged between 21 and 40 days were anesthetized with isoflurane, mounted in a stereotaxic frame, and a small craniotomy made above the target region. To achieve sparse labelling, a mixture of pENN‐AAV‐hSyn‐Cre‐WPRE‐hGH (Addgene # 105553‐AAV1) (titer ≥ 2 × 10⁹ vg mL⁻¹) and pAAV‐hsyn‐flex‐Voltron‐ST (Addgene # 119036‐AAV1) (titer ≥ 1 × 10¹² vg mL⁻¹) injected into the primary visual cortex (stereotaxic coordinates AP −3.8, ML 3.0, DV −0.3, and −0.6) and CA1 (AP −3.8, ML 3.0, DV −1.3). Injection coordinates were calculated relative to bregma and 200 nL of the virus mixture was injected at each coordinate. JF525‐HTL (100 nMol, Lavis Lab, Janelia Research Campus, HHMI) was mixed with 20 µL of DMSO (Sigma, D2650‐5 × 5 ML), 20 µL of Pluronic F‐127 (20% Solution in DMSO) (ThermoFischer Scientific P3000MP) and 80 µL of 1x PBS and administered intravenously into the lateral tail vein of mice 24–72 h prior to experimentation.

Tian T., Yuan Y., Mitra S., Gyongy I, & Nolan M.F. (2022). Single Photon Kilohertz Frame Rate Imaging of Neural Activity. Advanced Science, 9(31), 2203018.

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Producing Adeno-Associated Viral Vectors

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The following plasmids were purchased from Addgene: AAV-hSyn-eGFP (#114213), AAV-hSyn-hM3D(G_q)-mCherry (#50474), AAV-hSyn-DIO-hM3D(G_q)-mCherry (#44361), and pENN-AAV-hSyn-CRE-WPRE.hGH (#105553). AAV8-hSyn-DIO-hM3D(G_q)-eGFP-WPREs was purchased from BrainVTA. To produce viral particles, HEK293T cells were grown in DMEM (low glucose, GlutaMAX) supplemented with 10% fetal bovine serum. Cells were transfected by using polyethylenimine solution with a 1:1:1 molar ratio of helper, capsid, and desired construct plasmids in RPMI medium without glutamine and serum. 5 h after transfection, the media was changed back to serum-supplemented DMEM and the cells were kept for 3 d before harvesting. The supernatant was harvested 2 and 3 d after transfection and kept at 4°C. The supernatant was centrifuged (4,200 rpm for 10 min) to collect cell debris and then filtered (0.45 micron). For concentration, 15-min centrifugation rounds (4,200 rpm, 4°C) through 100 kD AMICON filters were performed. Titers were determined using qPCR. Viral aliquots were stored at −80°C until used.

Van Steenbergen V., Burattini L., Trumpp M., Fourneau J., Aljović A., Chahin M., Oh H., D’Ambra M, & Bareyre F.M. (2022). Coordinated neurostimulation promotes circuit rewiring and unlocks recovery after spinal cord injury. The Journal of Experimental Medicine, 220(3), e20220615.

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