KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Primary Antibodies | ||
| Cleaved Caspase-3 (Asp175) | Cell Signaling | 9661 |
| CTIP2 | Abcam | ab18465 |
| DCX | Santa Cruz | sc-8066 |
| GAD-65/67 | Santa Cruz | sc-365180 |
| GFP | Aves | GFP-1020 |
| KI67 | BD | 550609 |
| KIF26A | Sigma | HPA046882-100UL |
| KIF26A | Sigma | SAB1407288-50UG |
| KIF26A | Novus | NBP2-14158-25ul |
| mCherry | Abcam | ab205402 |
| Nestin | Aves | NES |
| NeuN | Millipore | MAB377 |
| Neuropilin-1 | R&D Systems | AF3870-SP |
| PAX6 | BD | 561664 |
| Phospho-FAK (Tyr576) | ThermoFisher | 700013 |
| Phospho-Histone H3 (Ser10) | Cell Signaling | 9706S |
| SATB2 | Abcam | ab51502 |
| SOX2 | Santa Cruz | sc-17320 |
| SOX2 | R&D Systems | AF2018 |
| TBR2 | Sigma | HPA028896-100UL |
| Acetylated tubulin | Sigma | T7451-100UL |
| βIII-tubulin | Abcam | ab18207 |
| Detyrosinated tubulin | Sigma | AB3201 |
| Chemicals, Peptides, and Recombinant Proteins | ||
| DMEM:F12 | Invitrogen | cat. # 11330032 |
| Neurobasal medium | Gibco | cat. # 21103049 |
| Phosphate Buffered Saline | Gibco | cat. # 10010023 |
| Non-essential Amino Acids | Gibco | cat. # 11140050 |
| Penicillin/Streptomycin | Gibco | cat. # 15140122 |
| 2-Mercaptoenthanol | Gibco | cat. # 21985023 |
| Glutamax | Gibco | cat. # 35050061 |
| Dorsomorphine | StemCell Technologies | cat. # 72102 |
| A83-01 | StemCell Technologies | cat. # 72022 |
| N2 Supplement | Gibco | cat. # 17502048 |
| B27 Supplements | Gibco | cat. # 17504044 |
| CHIR99021 | StemCell Technologies | cat. # 72052 |
| SB-431542 | StemCell Technologies | cat. # 72232 |
| Y-27632 | StemCell Technologies | cat. # 72304 |
| Nocodazole | StemCell Technologies | cat. # 74072 |
| GSK2256098 | Selleckchem | cat. # S8523 |
| Matrigel | Corning | cat. # 354230 |
| mTeSR™ Plus | StemCell Technologies | cat. # 100-0276 |
| ReLeSR | StemCell Technologies | cat. # 05872 |
| STEMdiff™ SMADi Neural Induction Kit | StemCell Technologies | cat. # 08582 |
| STEMdiff™ Forebrain Neuron Differentiation Kit | StemCell Technologies | cat. # 08600 |
| BrainPhys™ Neuronal Medium and SM1 Kit | StemCell Technologies | cat. # 05792 |
| Human Insulin solution | Sigma | cat. # I0516 |
| Ascorbic Acid | Sigma | cat. # 1043003 |
| Cyclic AMP | Sigma | cat. # A9501 |
| Paraformaldehyde (PFA) | Polysciences | cat. # 18814-10 |
| Sucrose | Sigma | cat. # S5016 |
| Tissue freezing medium | General Data | cat. # TFM-5 |
| Triton-X | Sigma | cat. # T9284 |
| donkey serum | Millipore | cat. # S30 |
| VECTASHIELD Vibrance Antifade Mounting Medium | Vector Laboratories | cat. # H-1700-10 |
| Retrievagen Antigen Retrieval Systems | Fisher | cat. # BDB550524 |
| RIPA Lysis and Extraction Buffer | ThermoFisher | cat. # 89901 |
| Anti-Adherence Rinsing Solution | StemCell Technologies | cat. # 07010 |
| Lipofectamine 3000 | ThermoFisher | cat. # L3000008 |
| Lipofectamine LTX | ThermoFisher | cat. # 15338030 |
| Poly-D-Lysine | ThermoFisher | cat. # A3890401 |
| Laminin Mouse Protein, Natural | ThermoFisher | cat. # 23017015 |
| Fast Green FCF | Sigma | cat. # F7252 |
| Precast Protein Gels | Bio-Rad | cat. # 4561093 |
| Mini PVDF Transfer Packs | Bio-Rad | cat. # 1704156EDU |
| Critical Commercial Assays | ||
| RNAscope™ Multiplex Fluorescent V2 Assay | ACD | cat. # 323110 |
| RNAscope® Probe- Hs-KIF26A-C1 | ACD | cat. # 856751 |
| RNAscope® Probe- Hs-RBFOX3-C3 | ACD | cat. # 415591-C3 |
| RNAscope® Probe- Hs-RORB-C2 | ACD | cat. # 446061-C2 |
| Click-iT® EdU Alexa Fluor® 488 Imaging kit | ThermoFisher | cat. # C10337 |
| ApopTag® Fluorescein In Situ Apoptosis Detection Kit | Millipore | cat. # S7110 |
| NovaSeq 6000 S1 Reagent Kit v1.5 (100 cycles) | Illumina | cat. # 20028319 |
| Chromium Next GEM Single Cell 3' Reagent Kit v3.1 | 10x Genomics | PN-1000121, PN-1000127 |
| Recombinant DNA and Viral Vectors | ||
| Lentivirus pLV[Exp]-mCherry-CMV>hKIF26A | VectorBuilder | VB200617-1157zbr |
| DNA plasmid pLV[Exp]-mCherry-CMV>hKIF26A[NM_015656.1] | VectorBuilder | VB200617-1157zbr |
| DNA plasmid pLV[Exp]-mCherry-CMV>{hKIF26A[NM_015656.1]*(R721C)} | VectorBuilder | VB210408-1104wqh |
| DNA plasmid pLV[Exp]-mCherry-CMV>{hKIF26A[NM_015656.1]*(P949S)} | VectorBuilder | VB210408-1092ysc |
| DNA plasmid pLV[Exp]-mCherry-CMV>{hKIF26A[NM_015656.1]*(A1559V)} | VectorBuilder | VB210408-1106ccf |
| DNA plasmid pLV[Exp]-mCherry-CMV>{hKIF26A[NM_015656.1]*R1624C} | VectorBuilder | VB220131-1120adp |
| DNA plasmid pRP[shRNA]-mCherry-U6>mKif26a[shRNA#1] | VectorBuilder | VB200226-1310hxy |
| DNA plasmid pRP[shRNA]-mCherry-U6>mKif26a[shRNA#2] | VectorBuilder | VB200226-1311htj |
| DNA plasmid pRP[shRNA]-mCherry-U6>mKif26a[shRNA#3] | VectorBuilder | VB200226-1312eag |
| DNA plasmid pLV[shRNA]-EGFP-U6>Scramble_shRNA#1 | VectorBuilder | VB010000-0001mty |
| DNA plasmid pLV[msgRNA]-mCherry-U6>Scramble[msgRNA#1] | VectorBuilder | VB200617-1158rnm |
| DNA plasmid pLV[shRNA]-EGFP-U6>hKIF26A[shRNA#1] | VectorBuilder | VB010000-0001mty |
| DNA plasmid pLV[shRNA]-EGFP-U6>hKIF26A[shRNA#2] | VectorBuilder | VB200817-1831kku |
| Deposited Data | ||
| Single cell RNA sequencing data of organoids | This paper | Accession code: XXX |
| Experimental Models: Cell Lines | ||
| Neuro-2a (N2A) mouse neuroblast cell line | ATCC | CCL-131 |
| HEK293T human kidney epithelial cell line | ATCC | CRL-3216 |
| SH-SY5Y human neuroblastoma epithelial cell line | ATCC | CRL-2266 |
| Human WT iPSC line “PGP1” derived from healthy male fibroblasts | Synthego (Lee et al., 2009) | PGP1 |
| Isogenic human iPSC line derived from “PGP1”, with frameshift mutation to knockout KIF26A gene using CRISPR-Cas9 editing | This paper | KO D17 |
| Isogenic human iPSC line derived from “PGP1”, with frameshift mutation to knockout KIF26A gene using CRISPR-Cas9 editing | This paper | KO N19 |
| Human WT iPSC line “280” derived from healthy male fibroblasts | Boston Children’s Hospital hESC Core (Schlaeger et al., 2015) | 280 |
| Isogenic human iPSC line derived from “280”, with frameshift mutation to knockout KIF26A gene using CRISPR-Cas9 editing | This paper | KO D2 |
| Isogenic human iPSC line derived from “280”, with frameshift mutation to knockout KIF26A gene using CRISPR-Cas9 editing | This paper | KO G4 |
| Experimental Models: Organisms/Strains | ||
| CD1 Mice | Charles River | Crl:CD1(ICR) |
| Ferrets | Marshall Bioresources | N/A |
| Software and Algorithms | ||
| Zeiss Zen (Blue edition) | Zeiss | https://www.zeiss.com/microscopy/us/products/microscope-software/zen.html |
| NextSeq System Suite v2.0 | Illumina | https://support.illumina.com/downloads/nextseq-system-suite-v2-0.html |
| ImageJ (Fiji) | NIH | https://imagej.nih.gov/ij/docs/guide/146-2.html |
| GraphPad Prism (8.0) | GraphPad Software Inc., La Jolla, CA, USA | https://www.graphpad.com/scientific-software/prism/ |
| Microsoft Excel | Microsoft | https://www.microsoft.com/en-us/p/excel/cfq7ttc0k7dx?activetab=pivot%3aoverviewtab |
| Adobe Photoshop (CC) | Adobe | https://www.adobe.com/products/photoshop.html |
| Adobe Illustrator (CC) | Adobe | https://www.adobe.com/products/illustrator.html |
| 10x Genomics Cell Ranger 4.0.0 and 5.0.0 | 10x Genomics | https://support.10xgenomics.com/single-cell-gene-expression/software/pipelines/latest/release-notes |
| TopHat (2.0) | Johns Hopkins University | https://ccb.jhu.edu/software/tophat/index.shtml |
| R, R studio with R package Seurat (v 2.3.4) | RStudio | https://cran.r-project.org/web/packages/Seurat/index.html |
| R studio with edgeR (v2.15) package | RStudio | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4023662/ |
| R package ClustifyR | RStudio | https://github.com/rnabioco/clustifyr |
| MathWorks Matlab | MathWorks | https://www.mathworks.com/ |
| MetaXpress | Molecular Devices | https://www.moleculardevices.com/products/cellular-imaging-systems/acquisition-and-analysis-software/metaxpress#gref |
| Other | ||
| AggreWell™400 | StemCell Technologies | cat. # 34425 |
| 12 Well glass bottom plate with high performance #1.5 cover glass | Cellvis | cat. # P12-1.5H-N |
| Costar® 6-well Ultra-Low Attachment Plate | Corning | cat. # 3471 |