KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| anti-TCAB1 rabbit polyclonal antibody | Proteintech | 14761-1-AP |
| anti-TCAB1 rabbit polyclonal antibody | Abcam | ab224444 |
| anti-TCAB1 rabbit polyclonal antibody | Novus Biologicals | NB100-68252 |
| anti-GAR1 rabbit polyclonal antibody | Proteintech | 11711-1-AP |
| anti-Coilin mouse monoclonal antibody | Abcam | ab87913 |
| anti-Dyskerin mouse monoclonal antibody | Santa Cruz Biotech. | sc-373956 |
| anti-TERT recombinant rabbit monoclonal antibody | Abcam | ab30202 |
| anti-TERT sheep polyclonal antibody | Scott Cohen | Abx120550 |
| anti-Fibrillarin mouse monoclonal antibody | Novus Biologicals | NB300-269 |
| anti-Lamin B1 rabbit polyclonal antibody | Proteintech | 12987-1-AP |
| anti-Rabbit Cy3 secondary antibody | Invitrogen | A-10520 |
| anti-Mouse Alexa 647 secondary antibody | Invitrogen | A-21235 |
| anti-Mouse Alexa 405 secondary antibody | Invitrogen | A-31553 |
| anti-Mouse HRP | Invitrogen | A-31430 |
| anti-Rabbit HRP | Invitrogen | A-31460 |
| Chemicals, peptides, and recombinant proteins | ||
| JF646 HaloTag Ligand | Grimm et al.35 | N/A |
| Prolong Diamond Antifade Mountant | Life Technologies | P36970 |
| Thymidine | Sigma | T1895 |
| Puromycin 10 mg/mL | Gibco | A1113803 |
| Lipofectamine 2000 | Invitrogen | 11668019 |
| dGTP [α-32P] 3000 Ci/mmol 10 mCi/mL | Perkin Elmer | BLU514H250UC |
| TERT peptide ARPAEEATSLEGALSGTRH | Cohen et al.36 | N/A |
| Dextrane sulfate | Sigma | D8906-10G |
| E. coli tRNA | Sigma | 10109541001 |
| Salmon Sperm DNA | Invitrogen | 15632011 |
| Vanadyl Ribonucleoside Complexes | Sigma | R3380-5ML |
| CHAPS | Fisher BioReagents | 211911 |
| SuperSignal West Femto Maximum Sensitivity Substrate | Thermo Scientific | 34096 |
| Clarity Western ECL substrate | Bio-Rad | 1705061 |
| Experimental models: Cell lines | ||
| HeLa EM2-11ht, TCAB1 KO | This paper | N/A |
| HeLa EM2-11ht, FLAG-HaloTag-TERT WT, HA-mEOS3.2-TRF2 | Schmidt et al.24 | N/A |
| HeLa EM2-11ht, FLAG-HaloTag-TERT WT, HA-mEOS3.2-TRF2, TCAB1 KO | This paper | N/A |
| HeLa EM2-11ht, FLAG-HaloTag-TERT WT, HA-mEOS3.2-TRF2, TR KO | This paper | N/A |
| HeLa EM2-11ht, FLAG-HaloTag-TERT WT, HA-mEOS3.2-TRF2, 3xMS2-TR | This paper | N/A |
| HeLa EM2-11ht, FLAG-HaloTag-TERT WT, HA-mEOS3.2-TRF2, 3xMS2-TR, TCAB1 KO | This paper | N/A |
| Oligonucleotides | ||
| All Oligonucleotides used in this study can be found in Table S1 | ||
| Recombinant DNA | ||
| pHTN HaloTag® CMV-neo | Promega | G7721 |
| pFastBac1 | Life Technologies | 10360014 |
| pX330 | Cong et al.37 | Addgene: 42230 |
| pmaxGFP | Lonza | Control from Kits |
| mCherry-TERT | Schmidt et al.38 | N/A |
| TERT (pVAN107) | Cristofari and Lingner39 | N/A |
| mCherry-Dyskerin | This paper | N/A |
| 3xFLAG-HaloTag-dyskerin | This paper | N/A |
| GFP-Nucleolin | Takagi et al.40 | Addgene: 28176 |
| GFP-NPM1 | Wang et al.41 | Addgene: 17578 |
| mTagBFP-Nucleus-7 (BFP-NLS) | Addgene | Addgene: 55265 |
| BFP-coilin | Schmidt et al.24 | N/A |
| pSUPER TR | Cristofari and Lingner39 | N/A |
| LhTRmin | Vogan et al.19 | N/A |
| pBS U3-hTR-500 | Fu and Collins42 | Addgene: 28170 |
| TR knockout HRD | This paper | N/A |
| pHAGE-UBC-MCP-mNeonGreen | This paper | N/A |
| 3xMS-TR-100-PURO HRD | This paper | N/A |
| Software and algorithms | ||
| MATLAB 2019a | Mathworks Inc., USA | http://mathworks.com |
| ParallelProcess_fastSPT_JF646.m | Hansen et al.27 | https://github.com/elifesciences-publications/SPT_LocAndTrack |
| ImageQuant TL 8.2 | Cytiva | cytivalifesciences.com |
| Spot-On | Hansen et al.27 | https://github.com/elifesciences-publications/spot-on-matlab |
| Other | ||
| SuperScript III | Invitrogen | 18080044 |
| Maxima SYBR Green qPCR Master Mix | Thermo Scientific | K0251 |
| Random Hexamers | Invitrogen | N8080127 |
| RiboLock RNase Inhibitor | Thermo Scientific | EO0382 |
| HOECHST 33342 | Invitrogen | 62249 |
| Protein G Agarose | Millipore Sigma | 11243233001 |
| 4-15% Mini-PROTEAN TGX precast protein gels | Bio-Rad | 4561083 |
| Trans-Blot Turbo RTA Mini 0.2 μm Nitrocellulose Kit | Bio-Rad | 1704270 |
| FxCycle Propidium iodide/RNase staining solution | Thermo Scientific | F10797 |
| RNeasy Mini Kit | Qiagen | 74106 |