Adeno-associated virus AAV9-Syn-FLEX-ChrimsonR-tdTomato, AAV2-hSyn-DIO-hm4D-mcherry, AAV9-hSyn-FLEX-tdTomato and AAV5-hSyn-Cre-eGFP were obtained from UNC Vector Core. AAVrg-CAG-GFP, AAV1-CB7-CI-TurboRFP, AAV1-CB7-CI-eGFP, AAVrg-Cre-eBFP and AAV1-hSyn1-SIO-stGtACR2-FusionRed were obtained from Addgene. All viral vectors were aliquoted and stored at −80 °C until used.
Aavrg cag gfp
Manufactured by Addgene
Sourced in United States
AAVrg-CAG-GFP is an adeno-associated virus (AAV) vector that contains the CAG promoter and the green fluorescent protein (GFP) gene. This vector can be used to express GFP in various cell types.
Automatically generated - may contain errors
Lab products found in correlation
Aav1 cb7 ci turborfp, by Addgene (2 mentions)
Aav1 cb7 ci egfp, by Addgene (2 mentions)
Aavrg cre ebfp, by Addgene (2 mentions)
Aav1 hsyn1 sio stgtacr2 fusionred, by Addgene (2 mentions)
Stereotaxic apparatus, by Stoelting (2 mentions)
Aavrg pmsyn1 ebfp cre, by Addgene (1 mentions)
Viral prep 51507 aavrg, by Addgene (1 mentions)
Aav dlx flex gfp, by Addgene (1 mentions)
Aav5 syn flex chrimsonr tdt, by Addgene (1 mentions)
Aavrg cag flex tdtomato, by Addgene (1 mentions)
7 protocols using aavrg cag gfp
1
Viral Vector Inventory Protocol
2
Viral Vector-Mediated Genetic Manipulation
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AAV8-hSyn-Cre (titer: 6.5 × 10¹² molecules/ml) was sourced from the UNC Vector Core (Chapel Hill, NC, USA). AAVrg-CAG-GFP (titer: 5 × 10¹² vg/mL, this construct was a gift from Edward Boyden to Addgene- viral prep # 37825-AAVrg) and AAVrg pmSyn1-EBFP-Cre (titer: 6 × 10¹² vg/mL, this construct was a gift from Hongkui Zeng to Addgene -viral prep # 51507-AAVrg; Madisen et al., 2015 (link)), were sourced from Addgene (MA, USA). All constructs were administered 1 ul bilaterally and a minimum of 3 weeks incubation was allowed.
For CaMKIIα cell-specific knockdown and fear conditioning and forced swim studies, 0.1 ul of AAV9.CamKII.HI.eGFP-Cre.WPRE.SV40 (CaMKIIα Cre) [Penn Vector Core] (titer: 6.544 × 1013 diluted to 6.544 × 1011] was injected bilaterally and allowed 5 weeks to incubate.
For CaMKIIα cell-specific knockdown and fear conditioning and forced swim studies, 0.1 ul of AAV9.CamKII.HI.eGFP-Cre.WPRE.SV40 (CaMKIIα Cre) [Penn Vector Core] (titer: 6.544 × 1013 diluted to 6.544 × 1011] was injected bilaterally and allowed 5 weeks to incubate.
3
Cre-Dependent AAV Viruses for Interneuron Targeting
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AAV viruses used in this study were as follows: AAV1.EF1a.DIO.hChR2(H134R)-eYFP.WPRE.hGH (UPenn AV-1–20298P), AAV1.hysn.hChR2(H134R)-eYFP.WPRE.hGH (UPenn AV-26973P), AAV1.EF1a.DIO.eYFP.WPRE.hGH (Upenn AV-1–27056), AAVrg.CAG.GFP (Addgene 37825-AAVrg), AAVrg.CAG.tdTomato (Addgene 59462-AAVrg). Additional viral constructs were assembled for Cre-dependent expression of a reporter under the control of the Dlx5/6 enhancer: AAV-Dlx-Flex-GFP (Addgene #83900) and AAV-Dlx-Flex-ChR2-mCherry (Dimidschstein et al., 2016 (link)). These constructs take advantage of the double-floxed inverted system, in which two consecutive and incompatible lox-sites are placed both in 5' and 3' of the reversed coding sequences of the viral reporter, restricting expression to Cre-expressing interneurons.
4
Versatile AAV Toolbox for Optogenetics
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All adeno-associated viruses (AAVs) were purchased from Addgene or custom-synthesized by the University of Pennsylvania vector core. The following AAVs were used: AAVrg-CAG-FLEX-tdTomato (Addgene 59462-AAVrg), AAVrg-CAG-GFP (Addgene: 37825-AAVrg), AAV5-pCAG-FLEX-EGFP (Addgene: 51502-AAV5), AAV9-EF1a-dfloxed-hChR2-EYFP (Addgene: 20298-AAV9), AAV5-Syn-FLEX-ChrimsonR-tdT (Addgene: 62723-AAV5), AAV9-EF1a-DIO-HA-SNAP-mGluR2-WPRE was previously reported.45 (link),48 (link) PORTL compound BGAG12,400 was synthesized by J. Broichhagen as previously described. 48 (link)
5
Stereotaxic Injection of AAVrg-CAG-GFP in Rat Cortex
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Rats were handled for two days prior to surgery. Rats were anaesthetized with ketamine (87 mg/kg) and xylazine (10 mg/kg), and they were placed in a stereotaxic apparatus (Stoelting, Woodale, IL) and received a unilateral injection of AAVrg-CAG-GFP (Addgene,30 (link)) into the IL (left- and right-side injections were counterbalanced). In order to perform the injection, a 22-gauge cannula was lowered into place (AP: + 3.00, ML: ± 0.6, DV: − 5.2). A 28-gauge internal cannula (connected to an infusion pump via PE 20 tubing) was inserted into the guide cannula and used to deliver 0.6 μL of the virus at a rate of 0.15 μL per minute and stayed in place for five minutes after the infusion was complete. After suturing rats were injected with Meloxicam (1 mg/kg) and they were returned to their home cages once ambulatory. Rats stayed in their home cage for about 7 weeks to allow for recovery and retrograde transport of the virus. Three rats died while under anesthesia, leaving 51 rats (94%) which successfully recovered from surgery.
6
Comprehensive AAV Viral Vector Toolkit
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Adeno-associated virus AAV9-Syn-FLEX-ChrimsonR-tdTomato, AAV2-hSyn-DIO-hm4D-mcherry, AAV9-hSyn-FLEX-tdTomato and AAV5-hSyn-Cre-eGFP were obtained from UNC Vector Core. AAVrg-CAG-GFP, AAV1-CB7-CI-TurboRFP, AAV1-CB7-CI-eGFP, AAVrg-Cre-eBFP and AAV1-hSyn1-SIO-stGtACR2-FusionRed were obtained from Addgene. All viral vectors were aliquoted and stored at -80°C until used.
7
AAV-mediated gene delivery to infralimbic cortex
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Rats were handled for two days prior to surgery. Rats were anaesthetized with ketamine (87 mg/kg) and xylazine (10 mg/kg), and they were placed in a stereotaxic apparatus (Stoelting, Woodale, IL) and received a unilateral injection of AAVrg-CAG-GFP (Addgene, [25] ) into the IL (left-and right-side injections were counterbalanced). In order to perform the injection, a 22-gauge cannula was lowered into place (AP: +3.00, ML: +/-0.6, DV: -5.2). A 28-gauge internal cannula (connected to an infusion pump via PE 20 tubing) was inserted into the guide cannula and used to deliver 0.6 μL of the virus at a rate of 0.15 μL per minute and stayed in place for five minutes after the infusion was complete. After suturing rats were injected with Meloxicam (1mg/kg) and they were returned to their home cages once ambulatory.
Rats stayed in their home cage for about 7 weeks to allow for recovery and retrograde transport of the virus. Three rats died while under anesthesia, leaving 51 rats (94%) which successfully recovered from surgery.
Rats stayed in their home cage for about 7 weeks to allow for recovery and retrograde transport of the virus. Three rats died while under anesthesia, leaving 51 rats (94%) which successfully recovered from surgery.
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