KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Biological samples | ||
| Smith-Lemli-Opitz patient-derived fibroblasts | Gift from Dr. Forbes Porter, NICHD | Refer to Table S3 |
| BJ human fibroblasts, foreskin derived | ATCC | CRL-2522 |
| Chemicals, peptides, and recombinant proteins | ||
| AY9944 | Cayman Chemical | Item No. 14611 |
| U-18666A | Cayman Chemical | Item No. 10009085 |
| Atorvastatin | Cayman Chemical | Item No. 10493 |
| Simvastatin | Cayman Chemical | Item No. 10010344 |
| methyl-β-cyclodextrin (MβCD)-cholesterol complex | Sigma | C4951 |
| Alfa Aesar MβCD (1303.31 738 g/mol) | Sigma Aldrich | AAJ6684714 |
| desmosterol | Avanti Polar Lipids | 700060 |
| lathosterol | Avanti Polar Lipids | 700069 |
| cholestenone | Avanti Polar Lipids | 700065 |
| cholesterol sulfate | Avanti Polar Lipids | 700016 |
| 4β-hydroxycholesterol | Avanti Polar Lipids | 700036 |
| 7-dehydrocholesterol (7DHC) | Avanti Polar Lipids | 70066 |
| cholesterol (ovine) | Avanti Polar Lipids | 700000 |
| coprostan-3-ol | Sigma Aldrich | C7578 |
| pyridine (CHROMASOLV Plus for HPLC) | Fischer Scientific | Cat#60-045-737 |
| latrunculin B | Calbiochem | Cat#428020 |
| N,O-bis(trimethylsilyl) trifluoroacetamide with 1% trimethylchlorosilane (BSTFA+ 1% TMCS) | Thermo Fisher | TS-38831 |
| Alexa Fluor™ 555 conjugated Transferrin | Thermo Fisher | T35352 |
| Electron Microscopy Sciences paraformaldehyde | Fisher Scientific | Cat#50-980-494 |
| glutaraldehyde, 8% aqueous solution (EM grade) | Electron Microscopy Sciences | 16020 |
| Corning human fibronectin | Fisher Scientific | Cat#CB-40008 |
| Critical commercial assays | ||
| CalPhos™ Mammalian Transfection kit | Clontech | Cat#631312 |
| Pierce MicroBCA Protein Assay | Thermo Scientific | Cat#23235 |
| 1,1’-Dioctadecyl-3,3,3′,3′-teramethylindocarboncyanine perchlorate (Dil) | Millipore Sigma | Cat#42364 |
| Experimental models: Cell lines | ||
| Human: HEK293T | ATCC | ACS-4500 |
| Human: HEK293T hCLTAEN-Tq2 | This paper | N/A |
| Human: SK-MEL-2 | ATCC | HTB-68 |
| Human: SK-MEL-2 hCLTAEN-Tq2 hDNM2EN-eGFP | Scott et al., 2018 | N/A |
| Oligonucleotides | ||
| CLTA gRNA: 5′-GCAGATGTAGTGTTTCCACA-3′ | Eurofins MWG | N/A |
| Recombinant DNA | ||
| Cas9 expression vector pX330-U6-Chimeric_BB-CBh-hSpCas9 | Cong et al., 2013 | Addgene plasmid #42230 |
| pDONOR2 hCLTA-Turquoise2-P2A-PuroR | Anderson et al., 2018 | N/A |
| Software and algorithms | ||
| GraphPad Prism 8.0.2 | https://www.graphpad.com/ | RRID:SCR_002798 |
| HCS Studio Cell Analysis software | https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cellular-imaging/hcs-hca.html | RRID:SCR_016787 |
| NIS-Elements analysis software | https://www.microscope.healthcare.nikon.com/products/software | RRID:SCR_014329 |
| ImageJ software (v1.8.0) | https://imagej.net/ | RRID:SCR_003070 |
| MassHunter (B.08.00) | https://www.agilent.com/en/product/software-informatics | RRID:SCR_015040 |
| MATLAB (2018a) | https://www.mathworks.com/ | RRID:SCR_001622 |
| MATLAB script: original CME analysis (cmeAnalysissoftware) | Aguet et al., 2013 | https://www.utsouthwestern.edu/labs/danuser/software/; https://github.com/DanuserLab/cmeAnalysis |
| Other | ||
| FACSJazz™ Cell Sorter | BD Biosciences | https://www.bdbiosciences.com/en-us/products/instruments/flow-cytometers/research-cell-sorters/bd-facsjazz |
| 7890A gas chromatography | Agilent Technologies | https://www.agilent.com/en/product/gas-chromatography |
| 5977B mass spectrometer | Agilent Technologies | https://www.agilent.com/en/product/gas-chromatography-massspectrometry-gc-ms |
| GC column: 0.18 mm ID × 20 m 1,4-bis(dimethylsiloxy)phenylene dimethyl polysiloxane | Restek | Cat#43602 |
| GC liners: 4 mm ID × 78.5 mm quartz wool liner | Restek | Cat#23309 |
| FluoroDish™ culture plates | World Precision Instruments Inc. | FD35–100 |
| Faramount Mounting Medium Dako | Aglient | 3307894 |