KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Bacterial and virus strains | ||
| E. coli Max Efficiency DH10B | Thermo Fisher | 18297010 |
| Chemicals, peptides, and recombinant proteins | ||
| GM1 species | This study | 10.17632/bk7cmygrdr.1 |
| Dimethylformamide (DMF) | Sigma Aldrich | 227056 |
| Dimethylsulfoxide (DMSO) | Sigma Aldrich | 472301 |
| Methanol | Fisher Scientific | A456–500 |
| Ethanol | Fisher Scientific | BP28184 |
| Acetonitrile | Fisher Scientific | A955–4 |
| TCEP | Sigma Aldrich | C4706 |
| soluble cholesterol complexed to MβCD | Sigma Aldrich | C4951 |
| defatted bovine serum albumin (dfBSA) | Sigma Aldrich | A8806 |
| fetal bovine serum depleted of HDL (HDL-) | Kalen Biomedical | 880100 |
| Simvastatin (statin) | Sigma Aldrich | S6196 |
| T0901317 | Sigma Aldrich | T2320 |
| U18666A | Tocris | 1638 |
| Filipin III | Sigma Aldrich | F4767 |
| Laurdan | Thermo Fisher | D250 |
| DMEM | Thermo Fisher | 12430112 |
| Opti-MEM | Thermo Fisher | 51985091 |
| Lipofectamine 2000 | Thermo Fisher | 11668030 |
| FASTDil™ | Thermo Fisher | D7756 |
| FASTDio™ | Thermo Fisher | D3898 |
| Alexa Fluor™ 488 Azide (also 568 and 647) | Thermo Fisher | A10266 |
| pHrodo™ Green Epidermal Growth Factor | Thermo Fisher | P35375 |
| pHrodo™ Red Transferrin Conjugate (also–AlexaFluor 488, 568, 647) | Thermo Fisher | P35376 |
| pHrodo™ iFL Green STP Ester | Thermo Fisher | P36012 |
| Dextran, Cascade Blue™, 3000 MW (also−488) | Thermo Fisher | D7132 |
| LysoTracker™ Red DND-99 (also DND-22) | Thermo Fisher | L7528 |
| Blasticidin | Thermo Fisher | A1113903 |
| Puromycin | Sigma-Aldrich | 540411 |
| Hygromycin | Thermo Fisher | 10687010 |
| Experimental models: Cell lines | ||
| A431 WT | ATCC | CRL-1555 |
| A431 SPTLC-2 KO | This study | N/A |
| A431 UGCG KO | This study | N/A |
| A431 CerS1 | This study | N/A |
| A431 CerS2 KO | This study | N/A |
| A431 CerS5 | This study | N/A |
| HT29 Cas9 | Pacheco et al. | Pacheco et al., 2018 |
| Hela WT | ATCC | CRM-CCL-2 |
| HEK293T | ATCC | ACS-4500 |
| CaCo BBE WT | ||
| SVGA Rab5-GFP | Chou et al. | Chou et al., 2016 |
| Oligonucleotides | ||
| SPTLC-2 ACAACTATCTTGGATTTGCA |
Pacheco et al. | Pacheco et al., 2018 |
| UGCG ACAACTATCTTGGATTTGCA |
Pacheco et al. | Pacheco et al., 2018 |
| CerS1 AGCGCTCTCGGGCATCTTGG GCTTCCCTGGAGCAGGTAGG GCAGCCGCCTACCTGCTCCA TACGCGCAGCTAGTGCAGCG |
This study | N/A |
| CerS2 GCTGCCTGTGAACTTGACCT TTCTTCATGTCATAGAACCA GGTCAGGTAGAAATGTTCCA GTAAAATGTGAATCTCCAGC |
This study | N/A |
| CerS5 GTATCCTGATAAGAAAAGGC TGATGAAACCAAGACCCAAG ATGCCAATACAGAGTGCACA ATAAAATGTGAATCTCCACC |
This study | N/A |
| Recombinant DNA | ||
| packaging plasmids pVSVg | Addgene | #8454 |
| packaging plasmids psPAX2 | Addgene | #12260 |
| lentiCas9-Blast plasmid | Addgene | #52962 |
| lentiGuide-Puro plasmid | Addgene | #52963 |
| dominant active RAB5Q79L-GFP plasmid | Addgene | #28046 |
| dominant active RAB5Q79L-GFP plasmid | Addgene | #35138 |
| Software and algorithms | ||
| ZENblack | Zeiss | https://www.zeiss.com/microscopy/us/products/microscope-software/zen.html |
| Fiji | https://fiji.sc/ | |
| Aivia 9 | Drvision Technologies | N/A |
| Matlab | MathWorks | https://www.mathworks.com/products/matlab.html |
| Graphpad Prism | Graphpad Prism | https://www.graphpad.com |
| Other | ||
| Lipidomics profiling | This study | Available upon request from lead contact |