Incubation:Article Title: Cell type-specific delivery by modular envelope design.
Article Snippet: .. After overnight incubation, media was exchanged Table 1 | Base constructs Name Source Comment Link psPAX2 Addgene plasmid # 12260 Gift from Didier Trono http://n2t.net/addgene:12260 MD2.G Addgene plasmid # 12259 Gift from Didier Trono http://n2t.net/addgene:12259 pCMV-15F11-HA-Halo Addgene plasmid # 129592 Gift from Tim Stasevich66 http://n2t.net/addgene:129592 RV-Cag-Dio-GFP Addgene plasmid # 87662 Gift from Oscar Marin67 http://n2t.net/addgene:87662 pME-puro-SNAP-FLAG-CD59 Addgene plasmid # 50374 Gift from Reika Watanabe68 http://n2t.net/addgene:50374 pHCMV-AmphoEnv Addgene plasmid # 15799 Gift from Miguel Sena-Esteves69 http://n2t.net/addgene:15799 pDisplay-mSA-EGFP-TM Addgene plasmid # 39863 Gift from Sheldon Park70 http://n2t.net/addgene:39863 pSLQ8703 pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1a-sfGFPT2A-rtta-WPRE Addgene plasmid # 140230 Gift from Stanley Qi71 http://n2t.net/addgene: 140230 m168 Addgene plasmid # 34886 Gift from Irvin Chen24 http://n2t.net/addgene:34886 MMLVgag-3NES-Cas9-NLS Addgene plasmid # 181752 Gift from David Liu6 http://n2t.net/addgene:181752 pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro Addgene plasmid # 141386 Gift from Nevan Krogan72 http://n2t.net/addgene:141386 pEx1-pEF-H2B-mCherry-T2A-rTetR-KRAB-Zeo Addgene plasmid # 78352 Gift from Michael Elowitz73 http://n2t.net/addgene:78352 lenti dCAS-VP64_Blast Addgene plasmid # 61425 74 http://n2t.net/addgene:61425 pLenti CMV rtTA3 Blast (w756-1) Addgene plasmid # 26429 Gift from Eric Campeau http://n2t.net/addgene:26429 plentiGuide-Puro Addgene plasmid # 52963 75 http://n2t.net/addgene:52963 Nature Communications | (2023) 14:5141 12 with fresh TCM (supplemented with 50U/ml rhIL-2) and cells were incubated for an additional 5 days with a media change at day 3 posttransduction. .. On day 6, cells were collected and half of the cells were used for analysis by flow cytometry, while the other half was used to stimulate T cell outgrowth by adding αCD3 and αCD28 T cell activator beads (Thermo Scientific Cat. # 11131D) in fresh T cell media supplemented with 50U/ml rhIL-2, 25U/ml rhIL-7 (PeproTech Cat. # 200-07), and 50U/ml rhIL-15 (PeproTech Cat. # 200-15).
Construct:Article Title: Cell type-specific delivery by modular envelope design.
Article Snippet: .. After overnight incubation, media was exchanged Table 1 | Base constructs Name Source Comment Link psPAX2 Addgene plasmid # 12260 Gift from Didier Trono http://n2t.net/addgene:12260 MD2.G Addgene plasmid # 12259 Gift from Didier Trono http://n2t.net/addgene:12259 pCMV-15F11-HA-Halo Addgene plasmid # 129592 Gift from Tim Stasevich66 http://n2t.net/addgene:129592 RV-Cag-Dio-GFP Addgene plasmid # 87662 Gift from Oscar Marin67 http://n2t.net/addgene:87662 pME-puro-SNAP-FLAG-CD59 Addgene plasmid # 50374 Gift from Reika Watanabe68 http://n2t.net/addgene:50374 pHCMV-AmphoEnv Addgene plasmid # 15799 Gift from Miguel Sena-Esteves69 http://n2t.net/addgene:15799 pDisplay-mSA-EGFP-TM Addgene plasmid # 39863 Gift from Sheldon Park70 http://n2t.net/addgene:39863 pSLQ8703 pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1a-sfGFPT2A-rtta-WPRE Addgene plasmid # 140230 Gift from Stanley Qi71 http://n2t.net/addgene: 140230 m168 Addgene plasmid # 34886 Gift from Irvin Chen24 http://n2t.net/addgene:34886 MMLVgag-3NES-Cas9-NLS Addgene plasmid # 181752 Gift from David Liu6 http://n2t.net/addgene:181752 pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro Addgene plasmid # 141386 Gift from Nevan Krogan72 http://n2t.net/addgene:141386 pEx1-pEF-H2B-mCherry-T2A-rTetR-KRAB-Zeo Addgene plasmid # 78352 Gift from Michael Elowitz73 http://n2t.net/addgene:78352 lenti dCAS-VP64_Blast Addgene plasmid # 61425 74 http://n2t.net/addgene:61425 pLenti CMV rtTA3 Blast (w756-1) Addgene plasmid # 26429 Gift from Eric Campeau http://n2t.net/addgene:26429 plentiGuide-Puro Addgene plasmid # 52963 75 http://n2t.net/addgene:52963 Nature Communications | (2023) 14:5141 12 with fresh TCM (supplemented with 50U/ml rhIL-2) and cells were incubated for an additional 5 days with a media change at day 3 posttransduction. .. On day 6, cells were collected and half of the cells were used for analysis by flow cytometry, while the other half was used to stimulate T cell outgrowth by adding αCD3 and αCD28 T cell activator beads (Thermo Scientific Cat. # 11131D) in fresh T cell media supplemented with 50U/ml rhIL-2, 25U/ml rhIL-7 (PeproTech Cat. # 200-07), and 50U/ml rhIL-15 (PeproTech Cat. # 200-15).
Plasmid Preparation:Article Title: Cell type-specific delivery by modular envelope design.
Article Snippet: .. After overnight incubation, media was exchanged Table 1 | Base constructs Name Source Comment Link psPAX2 Addgene plasmid # 12260 Gift from Didier Trono http://n2t.net/addgene:12260 MD2.G Addgene plasmid # 12259 Gift from Didier Trono http://n2t.net/addgene:12259 pCMV-15F11-HA-Halo Addgene plasmid # 129592 Gift from Tim Stasevich66 http://n2t.net/addgene:129592 RV-Cag-Dio-GFP Addgene plasmid # 87662 Gift from Oscar Marin67 http://n2t.net/addgene:87662 pME-puro-SNAP-FLAG-CD59 Addgene plasmid # 50374 Gift from Reika Watanabe68 http://n2t.net/addgene:50374 pHCMV-AmphoEnv Addgene plasmid # 15799 Gift from Miguel Sena-Esteves69 http://n2t.net/addgene:15799 pDisplay-mSA-EGFP-TM Addgene plasmid # 39863 Gift from Sheldon Park70 http://n2t.net/addgene:39863 pSLQ8703 pHR-R(TRE3G-Fluc-2XcrIFNg-SV40pA)-EF1a-sfGFPT2A-rtta-WPRE Addgene plasmid # 140230 Gift from Stanley Qi71 http://n2t.net/addgene: 140230 m168 Addgene plasmid # 34886 Gift from Irvin Chen24 http://n2t.net/addgene:34886 MMLVgag-3NES-Cas9-NLS Addgene plasmid # 181752 Gift from David Liu6 http://n2t.net/addgene:181752 pLVX-EF1alpha-SARS-CoV-2-M-2xStrep-IRES-Puro Addgene plasmid # 141386 Gift from Nevan Krogan72 http://n2t.net/addgene:141386 pEx1-pEF-H2B-mCherry-T2A-rTetR-KRAB-Zeo Addgene plasmid # 78352 Gift from Michael Elowitz73 http://n2t.net/addgene:78352 lenti dCAS-VP64_Blast Addgene plasmid # 61425 74 http://n2t.net/addgene:61425 pLenti CMV rtTA3 Blast (w756-1) Addgene plasmid # 26429 Gift from Eric Campeau http://n2t.net/addgene:26429 plentiGuide-Puro Addgene plasmid # 52963 75 http://n2t.net/addgene:52963 Nature Communications | (2023) 14:5141 12 with fresh TCM (supplemented with 50U/ml rhIL-2) and cells were incubated for an additional 5 days with a media change at day 3 posttransduction. .. On day 6, cells were collected and half of the cells were used for analysis by flow cytometry, while the other half was used to stimulate T cell outgrowth by adding αCD3 and αCD28 T cell activator beads (Thermo Scientific Cat. # 11131D) in fresh T cell media supplemented with 50U/ml rhIL-2, 25U/ml rhIL-7 (PeproTech Cat. # 200-07), and 50U/ml rhIL-15 (PeproTech Cat. # 200-15).
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