KEY RESOURCES TABLE.
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
|
| ||
| Antibodies | ||
|
| ||
| Goat anti-Sox2 | Santa Cruz Biotechnology | Cat# sc-17320; RRID:AB_2286684 |
| Rabbit anti-PTBP1 | Abcam | Cat# ab133734; RRID:AB_2814646 |
| Rabbit anti-RBPMS | Millipore | Cat# ABN1362; |
| Alexa Fluor 594 AffiniPure Donkey Anti-Mouse IgG (H + L) |
Jackson ImmunoResearch Labs | Cat# 715-585-151; RRID:AB_2340855 |
| Cy5 AffiniPure Donkey Anti-Rabbit IgG (H + L) |
Jackson ImmunoResearch Labs | Cat# 715-585-152; RRID:AB_2340607 |
|
| ||
| Chemicals, peptides, and recombinant proteins | ||
|
| ||
| Benzonase Nuclease | Sigma-Aldrich | Cat# E1024-25KU |
| DAPI | Thermo Fisher Scientific | Cat# 62248 |
| HEPES | Fisher BioReagents | Cat# BP310-100 |
| Iodixanol gradient | Alere Technologies | Prod.no.1114542 |
| L-Cysteine | MP Biochemicals | Cat# 101444 |
| N-Methyl-D-aspartic Acid (NMDA) | Millipore Sigma | Cat# 454575 |
| Papain | Worthington-Biochemical | Cat# LS003126 |
| Polyethylenimine (PEI) | Polysciences, Inc. | Cat# 23966-1 |
| Tamoxifen | Sigma-Aldrich | Cat# T5648 |
| TRIzol | Invitrogen | Cat# 15596026 |
| TURBO™ DNase | Invitrogen | Cat# AM2238 |
|
| ||
| Experimental models: Cell lines | ||
|
| ||
| AAVpro 293T Cell Line | Takara Bio | Cat# 632273 |
|
| ||
| Experimental models: Organisms/strains | ||
|
| ||
| Mouse: Glast-CreER: Tg(Slc1a3-cre/ERT)1Nat/J | The Jackson Laboratory | Cat# JAX:012586; RRID: IMSR_JAX:012586 |
| Mouse: R26R-EYFP: B6. 129X1-Gt(ROSA) 26Sortm1(EYFP)Cos/J | The Jackson Laboratory | Cat# JAX:006148; RRID: IMSR_JAX:006148 |
| Mouse: Rosa-CAG-LSL-Sun1-GFP: B6; 129-Gt(ROSA)26Sortm5(CAG–Sun1/sfGFP)Nat/J | The Jackson Laboratory | Cat# JAX:021039; RRID: IMSR_JAX:021039 |
| Mouse: Rosa-CAG-LSL-tdTomato: B6. Cg-Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J | The Jackson Laboratory | Cat# JAX:007909; RRID: IMSR_JAX:007909 |
|
| ||
| Oligonucleotides | ||
|
| ||
| β-actin qPCR primer forward: 5′-CTGTCCCTGTATGCCTCTGG-3′ |
Integrated DNA technologies | N/A |
| β-actin qPCR primer reverse: 5′-ATGTCACGCACGATTTCC-3′ |
Integrated DNA technologies | N/A |
| CasRx qPCR primer forward: 5′-AGGCTGGAGAAGATCGTGG-3′ |
Integrated DNA technologies | N/A |
| CasRx qPCR primer reverse: 5′-GGACTCGCCGAAGTACCTCT-3′ |
Integrated DNA technologies | N/A |
| Ptbp1 qPCR primer forward: 5′-AGAGGAGGCTGCCAACACTA-3′ |
Integrated DNA technologies | N/A |
| Ptbp1 qPCR primer reverse: 5′ -ATTGCCATTCCTGCATCCA-3′ |
Integrated DNA technologies | N/A |
| RFP qPCR primer forward: 5′-GGCATCCCCGACTTCTTTA-3′ |
Integrated DNA technologies | N/A |
| RFP qPCR primer reverse: 5′-AGCCGCGTGTTTTCTTCTG-3′ |
Integrated DNA technologies | N/A |
| tdTomato qPCR primer forward: 5′-GAGCAAGGGCGAGGAGGTCA-3′ |
Integrated DNA technologies | N/A |
| tdTomato qPCR primer reverse: 5′-GATGTCGGCGGGGTGCTTC-3′ |
Integrated DNA technologies | N/A |
|
| ||
| Recombinant DNA | ||
|
| ||
| pAAV-CMV-LSL-RFP | Qian et al. (2020) | N/A |
| pAAV-CMV-LSL-RFP-ShPtbp1 | Qian et al. (2020) | N/A |
| pAAV-GFAP-CasRx | Zhou et al. (2020) | Cat# 154000; RRID:Addgene_154000 |
| pAAV-GFAP-CasRx-Pfibp1 | Zhou et al. (2020) | Cat# 154001; RRID:Addgene_154001 |
| pAAV-GFAP-Cre | this study | N/A |
| pAAV-GFAP-tdTomato | this study | N/A |
| pAAV-GFAP-GFP | this study | N/A |
| pUCmini-iCAP-PHP.eB | Chan et al. (2017) | Cat# 103005; RRID:Addgene_103005 |
|
| ||
| Software and algorithms | ||
|
| ||
| Adobe Photoshop | Adobe | https://www.adobe.com/products/photoshop.html |
| GraphPad Prism 8 | GraphPad | https://www.graphpad.com/scientific-software/prism |
| ImageJ | Schindelin et al. (2012) | https://imagej.net/software/fiji |
| Blender 2.9 | Blender | https://www.blender.org |