Plasmids and reagents
| Reagent or resource . | Source . | Identifier . |
|---|---|---|
| Recombinant DNA | ||
| Plasmid: PB-TO-hNGN2 | Gift from iPSC Neurodegenerative Disease Initiative (iNDI) & Michael Ward | RRID:Addgene_172115 |
| Plasmid: PiggyBac transposase vector | Transposagen | N/A |
| Plasmid: PGK mScarlet-Synaptophysin | This paper | RRID:Addgene_206145 |
| Plasmid: PGK EGFP | This paper | RRID:Addgene_216110 |
| Plasmid: PGK EGFP-RAB3A-WT | This paper | RRID:Addgene_206146 |
| Plasmid: PGK EGFP-RAB3A-Q81L | This paper | RRID:Addgene_206147 |
| Plasmid: PGK EGFP-RAB3A-T86E | This paper | RRID:Addgene_206148 |
| Plasmid: PGK EGFP-RAB3A-T86A | This paper | RRID:Addgene_206149 |
| Plasmid: CMV HA-MADD | This paper | RRID:Addgene_206150 |
| Plasmid: CMV HA-RIM2 | This paper; GST-RIM2-RBD was a gift from Ruben Bierings (Erasmus University Medical Center) (Kat et al., 2021) | RRID:Addgene_206151 |
| Plasmid: CMV SNAP-synapsin | This paper | RRID:Addgene_206152 |
| Plasmid: pBI-BFP-NL1 | (Aiken and Holzbaur, 2024) | RRID:Addgene_206153 |
| Chemicals, peptides, and recombinant proteins | ||
| MLi-2 | Tocris | Cat# 5756 |
| DMSO | Sigma-Aldrich | Cat# D2650 |
| Matrigel growth factor reduced | Corning | Cat# 354230 |
| Essential 8 medium | Thermo Fisher Scientific | Cat# A1517001 |
| ReLeSR | Stemcell Technologies | Cat# 05872 |
| Accutase | Sigma-Aldrich | Cat# A6964 |
| ROCK inhibitor Y-27632 | Selleckchem | Cat# S1049 |
| Knockout serum replacement | Thermo Fisher Scientific | Cat# 10828010 |
| DMEM/F-12, HEPES | Thermo Fisher Scientific | Cat# 11330032 |
| N2 supplement | Thermo Fisher Scientific | Cat# 17502048 |
| Non-essential amino acids (NEAA) | Thermo Fisher Scientific | Cat# 11140050 |
| GlutaMAX | Thermo Fisher Scientific | Cat# 35050061 |
| Doxycycline | Sigma-Aldrich | Cat# D9891 |
| Poly-L-Ornithine | Sigma-Aldrich | Cat# P3655 |
| BrainPhys neuronal medium | Stemcell Technologies | Cat# 05790 |
| Laminin | Corning | Cat# 354232 |
| BDNF | PeproTech | Cat# 450-02 |
| NT-3 | PeproTech | Cat# 450-03 |
| B27 supplement | Thermo Fisher Scientific | Cat# 17504-044 |
| Lipofectamine stem transfection reagent | Thermo Fisher Scientific | Cat# STEM00003 |
| Microcystin-LR | Sigma-Aldrich | Cat# 475815 |
| Halt protease and phosphatase inhibitor cocktail | Thermo Fisher Scientific | Cat# 78442 |
| 5-Fluoro-2′-deoxyuridine | Sigma-Aldrich | Cat# F0503 |
| Uridine | Sigma-Aldrich | Cat# U3003 |
| DMEM (Dulbecco’s modified eagle’s medium) | Corning | Cat# MT10-013-CV |
| FuGENE 6 | Promega Corp | Cat# E2692 |
| GFP-trap magnetic particles M-270 | ChromoTek | Cat# gtd |
| GFP-trap magnetic agarose | ChromoTek | Cat# gtma |
| Lambda (λ) protein phosphatase | New England BioLabs | Cat# P0753S |
| Critical commercial assays | ||
| BCA protein assay kit | Thermo Fisher Scientific | Cat# 23225 |
| Plasmid maxi kit | QIAGEN | Cat# 12163 |
| Experimental models: Cell lines | ||
| Human: KOLF2.1J WT iPSCs | B. Skarnes (Jackson Laboratories, Connecticut) | RRID: CVCL_B5P3 |
| Human: KOLF2.1J LRRK2-R1441H iPSCs | B. Skarnes (Jackson Laboratories, Connecticut) | N/A |
| Human: KOLF2.1J PPM1H KO iPSCs | This paper | RRID: CVCL_C7TY |
| Human: HEK293T | ATCC | RRID: CVCL_0063 |
| Software and algorithms | ||
| FIJI (release 2.9.0) | NIH, USA | http://fiji.sc, RRID:SCR_002285 |
| Prism 9 | GraphPad | https://www.graphpad.com/scientific-software/prism/, RRID:SCR_002798 |
| RStudio: Integrated Development for R (2021.09.2 Build 382) | RStudio Team | http://www.rstudio.com/, RRID:SCR_000432 |
| R package: nlme | Pinheiro J, Bates D, R Core Team | http://CRAN.R-project.org/package=nlme, RRID:SCR_015655 |
| Matlab R2022a | MathWorks | https://www.mathworks.com/products/matlab.html, RRID:SCR_001622 |
| Volocity | PerkinElmer | https://www.perkinelmer.com, RRID:SCR_002668 |
| VisiView 5.0.0.24 | Visitron | https://www.visitron.de/products/visiviewr-software.html, RRID:SCR_022546 |
| LI-COR image studio | LI-COR | https://www.licor.com/bio/image-studio/, RRID:SCR_015795 |
| Adobe Illustrator 2022 | Adobe | https://www.adobe.com/products/illustrator.html, RRID:SCR_010279 |
| BioRender | BioRender | https://biorender.com/, RRID:SCR_018361 |
| Other | ||
| 35 mm #1.5 glass bottom imaging dishes | MatTek | Cat# P35G-1.5-20-C |
| ProLong gold antifade mountant | Thermo Fisher Scientific | Cat# P36930 |
| Reagent or resource . | Source . | Identifier . |
|---|---|---|
| Recombinant DNA | ||
| Plasmid: PB-TO-hNGN2 | Gift from iPSC Neurodegenerative Disease Initiative (iNDI) & Michael Ward | RRID:Addgene_172115 |
| Plasmid: PiggyBac transposase vector | Transposagen | N/A |
| Plasmid: PGK mScarlet-Synaptophysin | This paper | RRID:Addgene_206145 |
| Plasmid: PGK EGFP | This paper | RRID:Addgene_216110 |
| Plasmid: PGK EGFP-RAB3A-WT | This paper | RRID:Addgene_206146 |
| Plasmid: PGK EGFP-RAB3A-Q81L | This paper | RRID:Addgene_206147 |
| Plasmid: PGK EGFP-RAB3A-T86E | This paper | RRID:Addgene_206148 |
| Plasmid: PGK EGFP-RAB3A-T86A | This paper | RRID:Addgene_206149 |
| Plasmid: CMV HA-MADD | This paper | RRID:Addgene_206150 |
| Plasmid: CMV HA-RIM2 | This paper; GST-RIM2-RBD was a gift from Ruben Bierings (Erasmus University Medical Center) (Kat et al., 2021) | RRID:Addgene_206151 |
| Plasmid: CMV SNAP-synapsin | This paper | RRID:Addgene_206152 |
| Plasmid: pBI-BFP-NL1 | (Aiken and Holzbaur, 2024) | RRID:Addgene_206153 |
| Chemicals, peptides, and recombinant proteins | ||
| MLi-2 | Tocris | Cat# 5756 |
| DMSO | Sigma-Aldrich | Cat# D2650 |
| Matrigel growth factor reduced | Corning | Cat# 354230 |
| Essential 8 medium | Thermo Fisher Scientific | Cat# A1517001 |
| ReLeSR | Stemcell Technologies | Cat# 05872 |
| Accutase | Sigma-Aldrich | Cat# A6964 |
| ROCK inhibitor Y-27632 | Selleckchem | Cat# S1049 |
| Knockout serum replacement | Thermo Fisher Scientific | Cat# 10828010 |
| DMEM/F-12, HEPES | Thermo Fisher Scientific | Cat# 11330032 |
| N2 supplement | Thermo Fisher Scientific | Cat# 17502048 |
| Non-essential amino acids (NEAA) | Thermo Fisher Scientific | Cat# 11140050 |
| GlutaMAX | Thermo Fisher Scientific | Cat# 35050061 |
| Doxycycline | Sigma-Aldrich | Cat# D9891 |
| Poly-L-Ornithine | Sigma-Aldrich | Cat# P3655 |
| BrainPhys neuronal medium | Stemcell Technologies | Cat# 05790 |
| Laminin | Corning | Cat# 354232 |
| BDNF | PeproTech | Cat# 450-02 |
| NT-3 | PeproTech | Cat# 450-03 |
| B27 supplement | Thermo Fisher Scientific | Cat# 17504-044 |
| Lipofectamine stem transfection reagent | Thermo Fisher Scientific | Cat# STEM00003 |
| Microcystin-LR | Sigma-Aldrich | Cat# 475815 |
| Halt protease and phosphatase inhibitor cocktail | Thermo Fisher Scientific | Cat# 78442 |
| 5-Fluoro-2′-deoxyuridine | Sigma-Aldrich | Cat# F0503 |
| Uridine | Sigma-Aldrich | Cat# U3003 |
| DMEM (Dulbecco’s modified eagle’s medium) | Corning | Cat# MT10-013-CV |
| FuGENE 6 | Promega Corp | Cat# E2692 |
| GFP-trap magnetic particles M-270 | ChromoTek | Cat# gtd |
| GFP-trap magnetic agarose | ChromoTek | Cat# gtma |
| Lambda (λ) protein phosphatase | New England BioLabs | Cat# P0753S |
| Critical commercial assays | ||
| BCA protein assay kit | Thermo Fisher Scientific | Cat# 23225 |
| Plasmid maxi kit | QIAGEN | Cat# 12163 |
| Experimental models: Cell lines | ||
| Human: KOLF2.1J WT iPSCs | B. Skarnes (Jackson Laboratories, Connecticut) | RRID: CVCL_B5P3 |
| Human: KOLF2.1J LRRK2-R1441H iPSCs | B. Skarnes (Jackson Laboratories, Connecticut) | N/A |
| Human: KOLF2.1J PPM1H KO iPSCs | This paper | RRID: CVCL_C7TY |
| Human: HEK293T | ATCC | RRID: CVCL_0063 |
| Software and algorithms | ||
| FIJI (release 2.9.0) | NIH, USA | http://fiji.sc, RRID:SCR_002285 |
| Prism 9 | GraphPad | https://www.graphpad.com/scientific-software/prism/, RRID:SCR_002798 |
| RStudio: Integrated Development for R (2021.09.2 Build 382) | RStudio Team | http://www.rstudio.com/, RRID:SCR_000432 |
| R package: nlme | Pinheiro J, Bates D, R Core Team | http://CRAN.R-project.org/package=nlme, RRID:SCR_015655 |
| Matlab R2022a | MathWorks | https://www.mathworks.com/products/matlab.html, RRID:SCR_001622 |
| Volocity | PerkinElmer | https://www.perkinelmer.com, RRID:SCR_002668 |
| VisiView 5.0.0.24 | Visitron | https://www.visitron.de/products/visiviewr-software.html, RRID:SCR_022546 |
| LI-COR image studio | LI-COR | https://www.licor.com/bio/image-studio/, RRID:SCR_015795 |
| Adobe Illustrator 2022 | Adobe | https://www.adobe.com/products/illustrator.html, RRID:SCR_010279 |
| BioRender | BioRender | https://biorender.com/, RRID:SCR_018361 |
| Other | ||
| 35 mm #1.5 glass bottom imaging dishes | MatTek | Cat# P35G-1.5-20-C |
| ProLong gold antifade mountant | Thermo Fisher Scientific | Cat# P36930 |