Raav2 retro helper

Manufactured by Addgene

The RAAV2-retro helper is a plasmid vector used in the production of recombinant adeno-associated virus (rAAV) particles. It contains the necessary viral genes required for the packaging of rAAV2 viral particles.

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Lab products found in correlation

Paav hsyn hm3d gq mcherry, by Addgene (1 mentions) Paav ef1a double floxed hchr2 h134r eyfp wpre hghpa, by Addgene (1 mentions) Paav cag tdtomato, by Addgene (1 mentions) Pucmini icap php eb, by Addgene (1 mentions) Paav cag gfp, by Addgene (1 mentions) Paav hsyn dio hm3d gq mcherry, by Addgene (1 mentions) Paav hsyn dio hm4d gi mcherry, by Addgene (1 mentions) Paav hsyn egfp, by Addgene (1 mentions)

2 protocols using raav2 retro helper

Plasmid Purification and AAV Production

Cited 1 time

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Plasmids for transfection were purified by using Qiagen's endotoxin-free HiSpeed Gigaprep kit (Qiagen) and quantified by spectrophotometry. The integrity of the cis plasmids and ITRs were confirmed by restriction digest and NGS. Complete reagent information is listed in Supplementary Table S5.
The following plasmids were used to prepare the rAAV described in this study; pAAV-EF1a-double floxed-hChR2(H134R)-EYFP-WPRE-HGHpA (Addgene 20298, a gift of Karl Deisseroth, unpublished) AAV-Cre-GFP (Addgene 68544, a gift of Eric Nestler), AAV-EF1a-DIO-GCaMP6s-P2A-nls-dTomato (Addgene 51082, a gift of Jonathan Ting), AAV-FLEX-rev-ChR2(H134R)-mCherry (Addgene 18916, a gift from Scott Sternson), rAAV2-retro helper (Addgene 81070, a gift of Alla Karpova), pAAV-hSyn-DIO-hM3D(Gq)-mCherry (Addgene 44361, a gift of Bryan Roth), pAAV-hSyn-DIO-hM4D(Gi)-mCherry (Addgene 44362, a gift of Bryan Roth), pAAV-hSyn-hM3D(Gq)-mCherry (Addgene 50474, a gift of Bryan Roth, unpublished), pAAV-hSyn-EGFP(Addgene 50465, a gift of Bryan Roth, unpublished), pAAV-CAG-GFP (Addgene 37825, a gift from Edward Boyden, unpublished), pAAV-CAG-tdTomato (Addgene 59462, a gift of Edward Boyden, unpublished), and pUCmini-iCAP-PHP.eB (Addgene 103005, a gift of Viviana Gradinaru).^{15–20 (link)}

Guerin K., Rego M., Bourges D., Ersing I., Haery L., Harten DeMaio K., Sanders E., Tasissa M., Kostman M., Tillgren M., Makana Hanley L., Mueller I., Mitsopoulos A, & Fan M. (2020). A Novel Next-Generation Sequencing and Analysis Platform to Assess the Identity of Recombinant Adeno-Associated Viral Preparations from Viral DNA Extracts. Human Gene Therapy, 31(11-12), 664-678.

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Generating pAAV-U6-mCherry Construct

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To generate pAAV-U6-mCherry, the mCherry sequence was PCR amplified from pAAV-Ef1a-DIO-DSE-mCherry-PSE-shRNA. AvrII-WPRE-pA (purchased from Addgene; Cat #129669), the EGFP sequence was eliminated from pAAV-U6-EGFP by digestion with MluI and BsrGI, and the mCherry sequence was subcloned into the vector’s MluI/BsrGI enzyme site. The following plasmids were obtained commercially: rAAV2-retro helper (Addgene; Cat# 81070), pAAV-RAM-d2TTA::TRE-NLS-mKate2-WPREpA (Addgene; Cat# 84474), pAAV-hSynI-FLEX-TVA-P2A-EGFP-2A-oG (Addgene; Cat# 85225), EnvA G-deleted Rabies-mCherry (Addgene; Cat# 32626), and pAAV-EF1α-fDIO-hM4Di-mCherry (Addgene; Cat# 50461). pAAV-EF1α-fDIO-NLS-GFP-∆Cre and pAAV-EF1α-fDIO-NLS-GFP-Cre were constructed by PCR amplification of the NLS-∆Cre-GFP and NLS-Cre-GFP segment from pAAV-hSynI-NLS-∆Cre-GFP and pAAV-hSynI-NLS-Cre-GFP vector, respectively, followed by subcloning into the pAAV-EF1α-fDIO at BsrGI and NheI sites. The following plasmids were previously described: pAAV-U6-EGFP^{80 (link)}; pAAV-hSynI-∆Cre-EGFP and pAAV-hSynI-Cre-EGFP^{42 (link)} and pAAV-phSynI-WGA-IRES-mCherry-Flpo-bGHGpA^{81 (link)}; and pAAV-DIO-hM3Dq-2A-mCherry^{82 (link)}.

Han K.A., Yoon T.H., Kim J., Lee J., Lee J.Y., Jang G., Um J.W., Kim J.K, & Ko J. (2024). Specification of neural circuit architecture shaped by context-dependent patterned LAR-RPTP microexons. Nature Communications, 15, 1624.

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