Reagents and tools table.
| Reagent or resource | Source | Identifier |
|---|---|---|
| Antibodies | ||
| Mouse IgG1 anti-β-tubulin IV | Emergo Biogenex | #MU178-UC |
| Rabbit anti-β-tubulin IV | Abcam | #ab179509 |
| Rabbit anti-MUC5AC | Abcam | #ab198294 |
| Rabbit anti-P63 | Abcam | #ab124762 |
| Mouse IgG1 anti cytokeratin 5 | Abcam | #ab17130 |
| Mouse IgG1 anti-CC10 | Acris, Origene | #AM26360PU-N |
| Goat anti-Mouse IgG1, Alexa Fluor 488 | Invitrogen | #A-21121 |
| Goat anti-Mouse IgG1, Alexa Fluor 647 | Invitrogen | #A-21240 |
| Goat anti-Rabbit IgG, Alexa Fluor 488 | Invitrogen | #A-11034 |
| Goat anti-Rabbit IgG, Alexa Fluor 546 | Invitrogen | #A-11035 |
| Biological samples | ||
| Nasal brushings | UMC Utrecht | Protocol ID: 16/586, NL54885.041.16 |
| Nasal turbinate tissue | Academic Medical Center Amsterdam | N/A |
| Intestinal organoids | HUB | https://huborganoids.nl/ |
| Chemicals, peptides, and recombinant proteins | ||
| TrypLE express enzyme | Thermo Fisher Scientific | # 12605010 |
| Sputolysin | Calbiochem | #560000-10 |
| Collagen IV | Sigma-Aldrich | #C7521 |
| CryoStor CS10 | STEMCELL Technologies | #07930 |
| PureCol | Advanced BioMatrix | #5005 |
| Bronchial epithelial cell medium-basal (BEpiCM-b) | ScienCell | #3211 |
| Advanced DMEM F12 | Thermo Fisher Scientific | #12634-028 |
| B-27 Supplement, serum free | Thermo Fisher Scientific | #12587010 |
| GlutaMAX Supplement | Thermo Fisher Scientific | #35050-061 |
| HEPES | Thermo Fisher Scientific | #15630080 |
| (±)-Epinephrine hydrochloride | Sigma-Aldrich | #E4642 |
| Hydrocortisone | Sigma-Aldrich | #H0888 |
| 3,3′,5-Triiodo-L-thyronine sodium salt | Sigma-Aldrich | #T6397 |
| N-Acetyl-L-cysteine | Sigma-Aldrich | #A9165 |
| Nicotinamide | Sigma-Aldrich | #N0636 |
| SB02190 | Sigma-Aldrich | #S7067 |
| DMH-1 | Selleck Chemicals | #S7146 |
| A83-01 | Tocris | #2939/10 |
| Y-27632 | Selleck Chemicals | #S1049 |
| DAPT | Thermo Fisher Scientific | #15467109 |
| TTNPB | Cayman | #16144-1 |
| Recombinant human FGF-7 | PeproTech | #100-19 |
| Recombinant human FGF-10 | PeproTech | #100-26 |
| Recombinant human EGF | PeproTech | #AF-100-15 |
| Recombinant human HGF | PeproTech | #100-39H |
| Recombinant Neuregulin-1β | PeproTech | #100-03 |
| Recombinant Interleukin 1β | PeproTech | #200-01B |
| Penicillin–Streptomycin | Thermo Fisher Scientific | #15070-063 |
| Primocin | InvivoGen | #ant-pm-2 |
| Amphotericin B | Thermo Fisher Scientific | #15290018 |
| Gentamicin | Sigma-Aldrich | #G1397 |
| Vancomycin | Sigma-Aldrich | #SBR00001 |
| Collagenase type II | Thermo Fisher Scientific | #17101-015 |
| CFTR multi-guide sgRNA | Synthego | |
| 2NLS-Cas9 nuclease | Synthego | |
| OptiMEM | Thermo Fisher Scientific | #31985062 |
| CFTRinh-172 | Sigma-Aldrich | #C2992 |
| GlyH101 | Sigma-Aldrich | #219671 |
| VX-809 | Selleck Chemicals | #S1565 |
| VX-661 | Selleck Chemicals | #S7059 |
| VX-770 | Selleck Chemicals | #S1144 |
| Correctors C1-18 | Cystic Fibrosis Foundation Therapeutics | https://www.cff.org/Research/Researcher-Resources/Tools-and-Resources/CFTR-Chemical-Compound-Program/ |
| Forskolin | Sigma-Aldrich | #F3917 |
| 3-Isobutyl-1-methylxanthine | Sigma-Aldrich | #I5879 |
| Amiloride | Sigma-Aldrich | #1019701 |
| Calcein green acetoxymethyl (AM) | Invitrogen | #C34852 |
| Alexa Fluor 555 Phalloidin | Invitrogen | #A34055 |
| Phalloidin-iFluor 405 | Abcam | #ab176752 |
| ProLong Gold antifade reagent | Thermo Fisher Scientific | #P36934 |
| ProLong Gold antifade reagent with DAPI | Thermo Fisher Scientific | #P36935 |
| iQ SYBR Green Supermix | Bio-Rad | #1708880 |
| Critical Commercial Assays | ||
| RNeasy Mini Kit | QIAGEN | #74104 |
| iScript cDNA synthesis kit | Bio-Rad | #1708891 |
| Quick-DNA Microprep Kit | Zymo Research | #D3020 |
| GoTaq G2 Flexi DNA polymerase | Promega | #M7805 |
| Experimental Models: Cell Lines | ||
| Human nasal airway epithelial cells (HNEC) | This paper | N/A |
| Hek293T – R-spondin-1 mFc cell line | Trevigen Cat. no. 3710-001- | N/A |
| Oligonucleotides | ||
| Primers used for this manuscript | Table S5 | |
| Equipment | ||
| Zeiss LSM800 confocal microscopy | Zeiss | N/A |
| Leica SP8X confocal microscope | Leica | N/A |
| Leica THUNDER imager | Leica | N/A |
| Leica TCS SP8 STED 3X microscope | Leica | N/A |
| NEPA21 | NEPA | N/A |
| Ussing chamber system | Physiologic Instruments | N/A |
| Voltage clamp | World Precision Instruments | #DVC-1000 |
| PowerLab | AD Instruments | #8/30 |
| NanoDrop spectrophotometer | Thermo Fisher Scientific | N/A |
| CFX96 real-time detection machine | Bio-Rad | N/A |
| Software and Algorithms | ||
| Zen Blue Software | Zeiss | https://www.zeiss.com/microscopy/int/products/microscope-software/zen.html |
| Prism 8 | GraphPad Software Inc. | https://www.graphpad.com/scientific-software/prism/ |
| Microsoft Excel | Microsoft Corporation | https://office.microsoft.com/excel |
| Adobe Illustrator | Adobe | https://www.adobe.com/nl/products/illustrator.html |
| LabChart 6 | AD Instruments | https://labchart.software.informer.com/6.0/ |
| CFX Manager 3.1 | Bio-Rad | https://www.bio-rad.com/en-us/sku/1845000-cfx-manager-software?ID=1845000 |
| LAS X software | Leica | https://www.leica-microsystems.com/products/microscope-software/p/leica-las-x-ls/ |
| ImageJ/FIJI | https://imagej.net/Fiji/Downloads | |
| ICE analysis tool | Synthego | https://ice.synthego.com/#/ |
| Others | ||
| Cytological brush | CooperSurgical | #C0004 |
| 12-mm Transwell with 0.4-μm Pore Polyester Membrane Insert | Corning | #3460 |
| 6.5-mm Transwell with 0.4-μm Pore Polyester Membrane Insert | Corning | #3470 |
| Matrigel | Corning | #354230 |
| BC isolation and expansion medium | Table S2 | |
| Air–liquid interface differentiation medium | Table S3 | |
| Airway organoid culture medium | Table S4 |