KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| Rabbit anti-GAPDH (Clone D16H11) | Cell Signaling Technology | Cat# 5174; RRID: AB_10622025 |
| Rabbit anti-Vinculin (Clone E1E9V) | Cell Signaling Technology | Cat# 13901; RRID: AB_2728678 |
| Mouse anti-EGLN1 (Clone 1D5F2) | Proteintech | Cat# 66589–1-Ig; RRID: AB_2881949 |
| Rabbit anti-PHD3/EGLN3 | Proteintech | Cat# 18325–1-AP; RRID: AB_10640673 |
| Rabbit anti-AIF (Clone D39D2) | Cell Signaling Technology | Cat# 5318, RRID:AB_10829473 |
| Anti-IE1 | Zhu et al.71 | N/A |
| Anti-PP28 | Silva et al.72 | N/A |
| Anti-UL26 | Munger et al.73 | N/A |
| Bacterial and virus strains | ||
| AD169-GFP (BADwt BAC clone, UL131Repair) | Wang & Shenk74 | N/A |
| TB40/E-UL99-mCherry | O’Connor & Shenk75 | N/A |
| HSV-YFP | Etienne et al.76 | N/A |
| HCoV-OC43 | BEI Resources | Cat# NR-52282 |
| CRISPRi EGLN lentiviral constructs | This paper | N/A |
| Chemicals, peptides, and recombinant proteins | ||
| Dulbecco’s Modified Eagle Medium (DMEM) | Invitrogen | Cat# 11965118 |
| Fetal Bovine Serum (FBS) | Biowest | Cat# S1620, Lot# 208N21 |
| Penicillin-Streptomycin | Life Technologies | Cat# 15140–122 |
| Matrigel Basement Membrane Matrix | Corning | Cat# 354262 |
| Collagenase Type II | Sigma-Aldrich | Cat# C2-28-100mg |
| Adaptaquin | MedChemExpress | Cat# HY-101449 |
| Valganciclovir | MedChemExpress | Cat# HY-A0032A |
| Phosphonoacetic acid (PAA) | MedChemExpress | HY-128744 |
| Hoechst 33352 | Invitrogen | Cat# H3570 |
| Propidium Iodide | MedChemExpress | Cat# HY-D0815 |
| FuGENE Transfection Reagent | Promega | Cat# E2691 |
| Polybrene | Millipore | Cat# TR1003G |
| Puromycin | MP Biomedicals | Cat# 0210055225 |
| QuickExtract DNA Extraction Solution | Lugigen | N/A |
| Seahorse XF Calibrant | Agilent | Cat# 100840–000 |
| Seahorse XF DMEM Medium | Agilent | Cat# 103474–100 |
| GlutaMAX Supplement | Gibco | Cat# 35050–061 |
| Sodium Pyruvate (100x) | Gibco | Cat# 11360–070 |
| Glucose Solution | Gibco | Cat# A2494001 |
| PEG300 | Thermo Scientific | Cat# AC192220010 |
| Tween 80 | Thermo Scientific | Cat# AC278632500 |
| Critical commercial assays | ||
| Seahorse XF Cell Mito Stress Test | Agilent | Cat#103015–100 |
| Cell Fractionation Kit | Cell Signaling Technology | Cat# 9038 |
| Direct-zol RNA Miniprep Plus Kit | Zymo Research | Cat# R2072 |
| qScript cDNA Synthesis Kit | Quantabio | Cat# 101414–098 |
| Deposited data | ||
| Proteomics | This manuscript | ProteomeXchange: PXD074057 |
| Metabolomics | This manuscript | Metabolomics Workbench: PR002936, Project DOI:https://doi.org/10.21228/M85C3C |
| Experimental models: Cell lines | ||
| MRCT-hT (Telomerase-immortalized) | Cells from Shenk laboratory, telomerase transduction performed in Munger laboratory | N/A |
| Human Foreskin Fibroblasts (HFF) | Shenk laboratory | N/A |
| HEK293T | Shenk laboratory | N/A |
| ARPE19 epithelial cells | Shenk Laboratory | N/A |
| Experimental models: Organisms/strains | ||
| Mouse: NOD.Cg-Prkdc ^scid II2rg ^tm1Sug/JicTac (NOG-F) |
Taconic Biosciences | Model# NOG-F |
| Oligonucleotides | ||
| EGLN1 qPCR Primers | IDT | EGLN1 Forward: 5′-TGAGCAGCATGGACGACCTGAT-3′ EGLN1 Reverse: 5′-CGTACATAACCCGTTCCATTGCC-3′ |
| EGLN2 qPCR Primers | IDT | EGLN2 Forward: 5′-CTGTCTGGTATTTTGATGCCAAGG-3′ EGLN2 Reverse: 5′-CGGCTGTGATACAGGTACTTGG-3′ |
| EGLN3 qPCR Primers | IDT | EGLN3 Forward: 5′-GAACAGGTTATGTTCGCCACGTG-3′ EGLN2 Reverse: 5′-CCCTCTGGAAATATCCGCAGGA-3′ |
| IE1 qPCR Primers | IDT | IE1 Forward: 5′-CCATGTCCACTCGAACCTTAAT-3′ IE1 Reverse: 5′-TGAACAAGTGACCGAGGATTG-3′ |
| ND1 qPCR Primers | IDT | ND1 Forward: 5′-GGCTATATACAACTACGCAAAGGC-3′ ND1 Reverse: 5′-GGTAGATGTGGCGGGTTTTAGG-3′ |
| B-Actin qPCR Primers | IDT | B-Actin Forward: 5′-CACCATTGGCAATGAGCGGTTC-3′ B-Actin Reverse: 5′-AGGTCTTTGCGGATGTCCACGT-3′ |
| EGLN1 CRISPRi guide RNA | IDT | 5′-GGAAAGGACGAAACACCGGAGCTTAGGACTCGGAAGGTTTTAGAGCTAGAAATAGC-3′ |
| EGLN2 CRISPRi guide RNA | IDT | 5′-GGAAAGGACGAAACACCGCAGGGCGGCTGGCACAAAGTTTTAGAGCTAGAAATAGC-3′ |
| EGLN3 CRISPRi guide RNA | IDT | 5′-GGAAAGGACGAAACACCGCAGTGCGGCGCAAGGAGTGTTTTAGAGCTAGAAATAGC −3′ |
| Recombinant DNA | ||
| Plasmid: pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-puro |
Gift from Harris Lab | N/A |
| Plasmid: psPAX2 | Addgene | Cat# 12260 |
| Plasmid: pMD2.G | Addgene | Cat# 12259 |
| Software and algorithms | ||
| GraphPad Prism v10 | GraphPad Software | N/A |
| Gen5 v3.11 | Agilent (BioTek) | N/A |
| Proteome Discoverer v2.4 | Thermo Fisher Scientific | N/A |
| Compound Discover v3.3 | Thermo Fisher Scientific | N/A |
| El-MAVEN | Elucidata | N/A |
| MetaboAnalyst 6.0 | N/A | |
| Other | ||
| 384-well plate (black/clear bottom) | Corning | Cat# 3764 |
| Cytation 5 automated imaging reader | Agilent (BioTek) | N/A |