Key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Cell line (Homo sapiens) | HEK293T cells | Cell Culture Facility, UC Berkeley | N/A | |
| Cell line (H. sapiens) | U2OS cells | Cell Culture Facility, UC Berkeley | N/A | |
| Cell line (H. sapiens) | MDS-MB-231 cells | Cell Culture Facility, UC Berkeley | N/A | |
| Cell line (H. sapiens) | A549 cells | Cell Culture Facility, UC Berkeley | N/A | |
| Cell line (H. sapiens) | MCF7 cells | Cell Culture Facility, UC Berkeley | N/A | |
| Cell line (H. sapiens) | Hela cells | Cell Culture Facility, UC Berkeley | N/A | |
| Cell line (H. sapiens) | A431 cells | Cell Culture Facility, UC Berkeley | N/A | |
| Transfected construct (human) | siRNA to CLTC | QIAGEN | Hs_CLTC_10 FlexiTube siRNA, SI00299880 | Transfected construct (human) |
| Transfected construct (human) | siRNA to AP2B1 | QIAGEN | Hs_AP2B1_6 FlexiTube siRNA, SI02780085 | Transfected construct (human) |
| Transfected construct (human) | siRNA to CAV-1 | QIAGEN | Hs_CAV1_10 FlexiTube siRNA, SI00299642 | Transfected construct (human) |
| Transfected construct (human) | siRNA to FLOT2 | QIAGEN | Hs_FLOT2_5 FlexiTube siRNA, SI02781422 | Transfected construct (human) |
| Transfected construct (human) | siRNA to MFSD2A | QIAGEN | Hs_MFSD2_1 FlexiTube siRNA, SI04137854 | Transfected construct (human) |
| Antibody | Anti-GFP (rabbit polyclonal) | Torrey Pines Biolabs | Cat# AB_10013661 | WB (1:1000) |
| Antibody | Anti-myc (mouse monoclonal) | Cell Signaling Technology | Cat# 2276s | WB (1:1000) |
| Antibody | Anti-syntenin (rabbit polyclonal) | Santa Cruz Biotechnology | Cat# sc-48742 | WB (1:200) |
| Antibody | Anti-TSG101 (mouse monoclonal) | GeneTex | Cat# GTX70255 | WB (1:1000) |
| Antibody | Anti-CD81 (mouse monoclonal) | Santa Cruz Biotechnology | Cat# sc-166029 | WB (1:200) |
| Antibody | Anti-CD9 (rabbit monoclonal) | Cell Signaling Technology | Cat# AB_2798139 | WB (1:1000) |
| Antibody | Anti-CD63 (rabbit monoclonal) | Abcam | Cat# ab134045 | IF (1:300) |
| Antibody | Anti-CLTC (mouse monoclonal) | BD Transduction Laboratories | Cat# AB_397866 | WB (1:1000) |
| Antibody | Anti-AP2B1 (rabbit polyclonal) | Proteintech | Cat# 15690-1-AP | WB (1:1000) |
| Antibody | Anti-CAV1 (rabbit polyclonal) | Proteintech | Cat# 16447-1-AP | WB (1:1000) |
| Antibody | Anti-FLOT2 (mouse monoclonal) | BD Transduction Laboratories | Cat# 610384 | WB (1:1000) |
| Antibody | Anti-actin (rabbit monoclonal) | Cell Signaling Technology | Cat# 8457S | WB (1:1000) |
| Antibody | Anti-tubulin (mouse monoclonal) | Abcam | Cat# ab7291 | WB (1:1000) |
| Antibody | Anti-Arp2 (rabbit polyclonal) | Proteintech | Cat# 10922-1-AP | WB (1:1000) |
| Antibody | Anti-Arp3 (rabbit polyclonal) | Proteintech | Cat# 13822-1-AP | WB (1:1000) |
| Antibody | Anti-syncytin 1 (rabbit polyclonal) | Thermo Fisher Scientific | Cat# BS-2962R | WB (1:1000) |
| Antibody | Anti-syncytin 2 (rabbit polyclonal) | Thermo Fisher Scientific | Cat# PA5-109694 | WB (1:1000) |
| Antibody | Anti-MFSD2A (rabbit polyclonal) | OriGene Technologies | Cat# TA351394 | WB (1:1000) IF (1:300) |
| Antibody | Anti-Cas9 (mouse monoclonal) | Novus Biologicals | Cat# NBP2-36440 | WB (1:1000) |
| Recombinant DNA reagent | pUCOE- EF1a-dCas9-BFP-KRAB (plasmid) | Dr. Jonathan Weissman (Whitehead Institute, MIT) | Express dCas9-BFP-KRAB in cells for CRISPRi | |
| Recombinant DNA reagent | pMyc-streptavidin-CD63-mCherry (plasmid) | This study | Express Myc-streptavidin-CD63-mCherry in cells | |
| Recombinant DNA reagent | pSBP-flag-Cas9-GFP (plasmid) | This study | Express SBP-flag-Cas9-GFP in cells | |
| Recombinant DNA reagent | pHis-flag-Cas9-GFP (plasmid) | This study | His-flag-Cas9-GFP in cells | |
| Recombinant DNA reagent | pFluc-cas9 target-Nluc-IRES-mCherry (plasmid) | This study | Reporter for Cas9 editing | |
| Recombinant DNA reagent | pFluc-cas9 target-Nluc1-Nluc2-IRES-mCherry (plasmid) | This study | Reporter for Cas9 editing | |
| Recombinant DNA reagent | pRluc-FLXXUC-IRES-mCherry (plasmid) | This study | Reporter for Cas9 editing | |
| Recombinant DNA reagent | pLKO.1-shactin (plasmid) | This study | Lentiviral construct to transfect and express the shRNA | |
| Recombinant DNA reagent | pLKO.1-sharp2 (plasmid) | This study | Lentiviral construct to transfect and express the shRNA | |
| Recombinant DNA reagent | pLKO.1-sharp3 (plasmid) | This study | Lentiviral construct to transfect and express the shRNA | |
| Recombinant DNA reagent | pCD63-7GFP11-IRES-CFP (plasmid) | This study | Express CD63-7GFP11 and CFP in cells | |
| Recombinant DNA reagent | pGFP10-IRES-mCherry (plasmid) | This study | Express GFP10 and mCherry in cells | |
| Recombinant DNA reagent | pU6_sgRNA_CAG_puroR (plasmid) | Addgene | Express sgRNA for CRISPRi | |
| Recombinant DNA reagent | pSyncytin A-GFP (plasmid) | This study | Express Syncytin A-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A (1-416)-GFP (plasmid) | This study | Express Syncytin A (1-416)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A (1-445)-GFP (plasmid) | This study | Express Syncytin A (1-445)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A (1-493)-GFP (plasmid) | This study | Express Syncytin A (1-493)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A (1-531)-GFP (plasmid) | This study | Express Syncytin A (1-531)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A (417-617)-GFP (plasmid) | This study | Express Syncytin A (417-617)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A delete (417-536)-GFP (plasmid) | This study | Express Syncytin A delete (417-536)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A delete (445-536)-GFP (plasmid) | This study | Express Syncytin A delete (445-536)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin A delete (493-536)-GFP (plasmid) | This study | Express Syncytin A delete (493-536)-GFP in cells | |
| Recombinant DNA reagent | pSyncytin-1-BFP (plasmid) | This study | Syncytin-1-BFP in cells | |
| Recombinant DNA reagent | pSyncytin-2-BFP (plasmid) | This study | Syncytin-2-BFP in cells | |
| Recombinant DNA reagent | pLenti-Syncytin-1-GFP11 (plasmid) | This study | Express Syncytin-1-GFP11 in cells | |
| Recombinant DNA reagent | pLenti-Syncytin-2-GFP11 (plasmid) | This study | Express Syncytin-2-GFP11 in cells | |
| Recombinant DNA reagent | pSyncytin-1-GFP (plasmid) | This study | Express Syncytin-1-GFP in cells | |
| Recombinant DNA reagent | pSyncytin-2-GFP (plasmid) | This study | Express Syncytin-2-GFP in cells | |
| Recombinant DNA reagent | pMFSD2A-GFP (plasmid) | This study | Express MFSD2A-GFP in cells | |
| Chemical compound, drug | Biotin | Sigma | Cat# B4639 | |
| Chemical compound, drug | Proteinase K | Sigma | Cat# P2308 | |
| Chemical compound, drug | Micrococcal Nuclease | NEB | Cat# M0247S | |
| Chemical compound, drug | ANTI-FLAG M2 Affinity Gel | Sigma | Cat# A2220 | |
| Chemical compound, drug | SuperSignal West Pico PLUS | Thermo Fisher Scientific | Cat# PI34580 | |
| Chemical compound, drug | SuperSignal West Femto | Thermo Fisher Scientific | Cat# PI34096 | |
| Chemical compound, drug | TGIRT-III Enzyme | InGex | Matthew et al., PNAS (2017) | |
| Commercial assay or kit | Direct-zol RNA Miniprep Plus Kit | Zymo research | Cat# R2072 | |
| Commercial assay or kit | mirVana miRNA isolation kit | Thermo Fisher Scientific | Cat# AM1560 | |
| Commercial assay or kit | Nano-Glo Luciferase Assay | Promega | Cat# N1150 | |
| Commercial assay or kit | Luciferase Assay System | Promega | Cat# E4550 | |
| Commercial assay or kit | Luciferase reporter assay | Promega | Cat# E1910 | |
| Chemical compound, drug | PowerUp SYBR Green Master Mix | Thermo Fisher Scientific | Cat# A25741 | |
| Chemical compound, drug | Chlorpromazine | Sigma | Cat# C8138 | 5 µg/ml |
| Chemical compound, drug | LY294002 | Sigma | Cat# L9908 | 10 µM |
| Chemical compound, drug | Wortmannin | Sigma | Cat# W1628 | 1 µM |
| Chemical compound, drug | Latrunculin A | Enzo | Cat# BML-T119-0100 | 40, 80, 200 nM |
| Chemical compound, drug | Latrunculin B | Sigma | Cat# 428020 | 1, 2.5, 5 µM |
| Chemical compound, drug | SMIFH2 | Sigma | Cat# 344092 | 10, 25 µM |
| Chemical compound, drug | Hoechst 33342 | Thermo Fisher Scientific | Cat# H3570 | |
| Chemical compound, drug | Phrodo Red Transferrin Conjugate | Thermo Fisher Scientific | Cat# P35376 | |
| Chemical compound, drug | Phrodo Green Zymosan Bioparticles | Thermo Fisher Scientific | Cat# P35365 | |
| Chemical compound, drug | Molecular Probes pHrodo Green Dextran | Thermo Fisher Scientific | Cat# P35368 | |
| Chemical compound, drug | Sulfo-NHS-LC-Biotin | Thermo Fisher Scientific | Cat# 21335 | |
| Chemical compound, drug | GFP-Trap agarose beads | ChromoTek | Cat# gta-20 | |
| Chemical compound, drug | Streptavidin-HRP Conjugate | Thermo Fisher Scientific | Cat# SA10001 | |
| Chemical compound, drug | CF640R Wheat Germ Agglutinin (WGA) Conjugates | Biotium | Cat# 29026-1 | |
| Chemical compound, drug | Lipofectamine 2000 Transfection Reagent | Thermo Fisher Scientific | Cat# 11668019 | |
| Chemical compound, drug | Lipofectamine RNAiMAX Transfection Reagent | Thermo Fisher Scientific | Cat# 13778150 | |
| Other | Glass-bottom dish | Thermo Fisher Scientific | Cat# NC0699576 (Cellvis D35-20-1-N) | For confocal microscopy experiments |
| Other | Gridded glass-bottom dish | Thermo Fisher Scientific | Cat# NC1144968 | For CLEM experiments |
| Software, algorithm | Fiji | NIH | https://fiji.sc/ | |
| Software, algorithm | GraphPad Prism | GraphPad | https://www.graphpad.com | |
| Software, algorithm | Imaris | Oxford Instruments | https://imaris.oxinst.com/ |