Appendix 1—key resources table.
| Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (Homo sapiens) | MPI | GenBank | Gene ID: 4351 | |
| Genetic reagent (H. sapiens) | MPI-KO HT1080 | This paper | Clone #2, #3, which can be obtained from the Department of Glyco-Oncology and Medical Biochemistry, Osaka International Cancer Institute | |
| Genetic reagent (H. sapiens) | Fucci MPI-KO HT1080 | This paper | Clone #3 subclone (9-5), which can be obtained from the Department of Glyco-Oncology and Medical Biochemistry, Osaka International Cancer Institute | |
| Genetic reagent (H. sapiens) | MPI-KO HeLa | This paper | Clone #1, #2, which can be obtained from the Department of Glyco-Oncology and Medical Biochemistry, Osaka International Cancer Institute | |
| Genetic reagent (Mus musculus) | Wild-type mouse embryonic fibroblasts | DeRossi et al., 2006 | ||
| Genetic reagent (M. musculus) | MPI-KO mouse embryonic fibroblasts | DeRossi et al., 2006 | ||
| Cell line (H. sapiens) | HT1080 | Our laboratory stock | Validated by short tandem repeat profiling | |
| Cell line (H. sapiens) | HeLa | RIKEN BioResource Research Center | RCB0007 | |
| Cell line (H. sapiens) | KP-4 | RIKEN BioResource Research Center | RCB1005 | |
| Cell line (H. sapiens) | SK-OV-3 | American Type Culture Collection (ATCC) | HTB-77 | |
| Cell line (H. sapiens) | A549 | ATCC | CRM-CCL-185 | |
| Cell line (M. musculus) | MCA205 | A gift from Rosenberg lab | ||
| Cell line (H. sapiens) | The Plat-A retroviral packaging cell line | Cell Biolabs | RV-102 | |
| Antibody | Anti-MPI (rabbit polyclonal) | GeneTex | GTX103682 | WB (1:5000) |
| Antibody | Anti-β-actin (mouse monoclonal) | FUJIFILM Wako | 010-27841 | WB (1:10,000) |
| Antibody | Anti-H2AX (mouse monoclonal) | Santa Cruz | 938CT5.1.1 | WB (1:2000) |
| Antibody | Anti-phospho-H2AX (Ser139) (mouse monoclonal) | Millipore | JBW301 | WB (1:5000) Chromatin FACS (1:200) |
| Antibody | Anti-RPA2 (mouse monoclonal) | Santa Cruz | 9H8 | WB (1:5000) |
| Antibody | Anti-Phospho-RPA2 (S33) (rabbit polyclonal) | Bethyl | A300-246A | WB (1:10,000) |
| Antibody | Anti-MCM2 (rabbit monoclonal) | Cell Signaling Technology | D7G11 | WB (1:10,000) Chromatin FACS (1:200) |
| Antibody | Anti-MCM3 (mouse monoclonal) | Santa Cruz | E-8 | WB (1:10,000) |
| Antibody | Anti-MCM4 (rabbit polyclonal) | GeneTex | GTX109740 | WB (1:5000) |
| Antibody | Anti-MCM5 (rabbit polyclonal) | GeneTex | GTX33310 | WB (1:5000) |
| Antibody | Anti-MCM6 (mousemonoclonal) | Santa Cruz | H-8 | WB (1:20,000) |
| Antibody | Anti-MCM7 (mouse monoclonal) | Santa Cruz | 141.2 | WB (1:5000) |
| Antibody | Anti-CDT1 (rabbit monoclonal) | Abcam | ab202067 | WB (1:10,000) |
| Antibody | Anti-CDC6 (rabbit monoclonal) | Abcam | ab109315 | WB (1:1000) |
| Antibody | Anti-ORC2 (rat monoclonal) | Santa Cruz | 3G6 | WB (1:10,000) |
| Antibody | Anti-CDC45 (rabbit monoclonal) | Cell Signaling Technology | D7G6 | WB (1:10,000) |
| Antibody | Alexa Fluor 488-conjugated anti-BrdU antibody (mouse monoclonal) | BioLegend | 3D4 | FACS (1:100) |
| Antibody | Anti-rabbit IgG-Alexa Fluor 488 (donkey polyclonal) | Thermo Fisher Scientific | R37118 | Chromatin FACS (1:1000) |
| Antibody | Anti-mouse IgG-Alexa Fluor 647 (donkey polyclonal) | Thermo Fisher Scientific | A-31571 | Chromatin FACS (1:1000) |
| Recombinant DNA reagent | pENTR-hMPI | This paper | This reagent can be obtained from the Department of Glyco-Oncology and Medical Biochemistry, Osaka International Cancer Institute | |
| Recombinant DNA reagent | pMXs-Neo-hMPI | This paper | This reagent can be obtained from the Department of Cancer Drug Discovery and Development, Osaka International Cancer Institute | |
| Recombinant DNA reagent | mCherry-hCdt1(1/100)Cy(−)/pcDNA3 | RIKEN BioResource Research Center | RDB15459 | |
| Recombinant DNA reagent | pMXs-Neo to yield pMXs-Neo-mCherry-hCdt1(1/100)Cy(−) | This paper | This reagent can be obtained from the Department of Cancer Drug Discovery and Development, Osaka International Cancer Institute | |
| Recombinant DNA reagent | pMMLV-mVenus-hGem(1/110):IRES:Bsd | Vector Builder | ||
| Recombinant DNA reagent | Edit-R SMART Cas9_mCMV_(PuroR) expression plasmid | Dharmacon | U-005200-120 | |
| Recombinant DNA reagent | pENTR/D-TOPO vector | Thermo Fisher Scientific | K240020 | |
| Recombinant DNA reagent | pMXs-Neo retroviral expression vector | Cell Biolabs | RTV-011 | |
| Sequence-based reagent | Synthetic CRISPR RNAs for human MPI | Dharmacon | CM-011729-01 CM-011729-03 |
|
| Sequence-based reagent | trans-activating CRISPR RNA | Dharmacon | U-002005-05 | |
| Commercial assay or kit | VECTASTAIN ABC-HRP kit | Vector Laboratories | PK-4000 | |
| Commercial assay or kit | CellTiter-Glo 2.0 | Promega | G924B | |
| Commercial assay or kit | Cytotoxicity LDH Assay Kit-WST | Dojindo | CK12 | |
| Commercial assay or kit | PAGE Clean Up Kit | Nacalai | 06441-50 | |
| Commercial assay or kit | In-Fusion HD Cloning Kit | Clontech | 639648 | |
| Commercial assay or kit | RNeasy Mini Kit | QIAGEN | 74106 | |
| Commercial assay or kit | NucleoSpin Gel and PCR Clean-Up Kit | Clontech | 740609 | |
| Commercial assay or kit | THUNDERBIRD Next SYBR qPCR Mix | TOYOBO | QPX-201 | |
| Chemical compound, drug | Cellstain Hoechst 33342 solution | Dojindo | H342 | FACS (1:100) Chromatin FACS (1:200) |
| Chemical compound, drug | 5-Bromo-2’-deoxyuridine | Tokyo Chemical Industry | B1575 | |
| Chemical compound, drug | 2’-Deoxyadenosine | Wako | 046-18693 | |
| Chemical compound, drug | Thymidine | Wako | 203-19423 | |
| Chemical compound, drug | 2’-Deoxyguanosine | Tokyo Chemical Industry | D0052 | |
| Chemical compound, drug | 2’-Deoxycytidine | Tokyo Chemical Industry | D3583 | |
| Chemical compound, drug | Dialyzed FBS | Cytiva | SH30079.02 | |
| Chemical compound, drug | Cisplatin | Tokyo Chemical Industry | D3371 | |
| Chemical compound, drug | Doxorubicin | FUJIFILM Wako | 040-21521 | |
| Chemical compound, drug | IACS-010759 | Selleckchem | S8731 | |
| Chemical compound, drug | VE-821 | Sigma-Aldrich | SML1415 | |
| Chemical compound, drug | Hydroxyurea | FUJIFILM Wako | 085-06653 | |
| Chemical compound, drug | [13C6]-Glucose | Cambridge Isotope Laboratories | CLM-1396-1 | |
| Chemical compound, drug | cOmplete Protease Inhibitor Cocktail | Sigma-Aldrich | 11836170001 | |
| Chemical compound, drug | PhosSTOP | Sigma-Aldrich | 4906845001 | |
| Chemical compound, drug | Biotinylated concanavalin A | Cosmo Bio | J203 | Lectin blot (1:10,000) |
| Chemical compound, drug | TRIzol Reagent | Thermo Fisher Scientific | 15596026 | |
| Chemical compound, drug | Lipofectamine 3000 | Thermo Fisher Scientific | L3000015 | |
| Chemical compound, drug | Lipofectamine 2000 | Thermo Fisher Scientific | 11668027 | |
| Software, algorithm | MASCOT 2.8 | Matrix Science | ||
| Software, algorithm | Proteome Discoverer 2.4 | Thermo Fisher Scientific | ||
| Software, algorithm | ImageJ | Schneider et al., 2012 | ||
| Software, algorithm | Trackmate | Tinevez et al., 2017 | ||
| Software, algorithm | MATLAB | MathWorks |