Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| cell line (Homo sapiens, Male) |
HCT116 cells | Other | RRID:CVCL_0291 | Prescott Deininger at Tulane Univeristy LCRC |
| cell line (H. sapiens, Male) |
HCT116 + Mlh1 | This paper | NA | HCT116 cells stably expressing human Mlh1-ORF via lentivirus-mediated integration |
| cell line (H. sapiens, Male) |
Exo-; Exonuclease-deficient HCT116 Cells |
This paper | NA | HCT116 cells infected with rAAV containing D275A and E277A POLE mutations |
| cell line (H. sapiens, Male) |
Exo-; Exonuclease-deficient HCT116 Cells + Mlh1 |
This paper | NA | HCT116 cells stably expressing human Mlh1-ORF via lentivirus- mediated integration and infected with rAAV containing D275A and E277A POLE mutations |
| recombinant DNA reagent |
ExoI-targeting rAAV vector | This paper | NA | Homology arms/SEPT Cassette/Exo- mutations |
| recombinant DNA reagent |
pCMV-XL5-Mlh1 | Other | NA | Victoria Belancio at Tulane Univeristy LCRC |
| antibody | Mlh1 Antibody | Pharmingen | G168-728; RRID: AB_395227 |
Rabbit monoclonal; (1:100) in Milk (1%) TBST (1X) x 1 hr at RT |
| chemical compound, drug | 6-Thioguanine; 6-TG | Sigma-Aldrich | A4882 | Used at 5 ug/mL final concentration |
| chemical compound, drug | Hypoxanthine-Aminopterin- Thymidine; HAT |
Thermo Fisher Scientific | 21060017 | Used at 1X final concentration |
| chemical compound, drug | Geneticin; G418 | Thermo Fisher Scientific | 10131027 | Used at 400 ug/mL final concentration |
| other | Ad-CMV-Cre | Vector Biolabs | 1045 | Adenovirus expressing Cre recombinase for excision of SEPT cassette from ExoI-targeted cell lines |
| software, algorithm | BWA-MEM v0.7.8 | PMID: 19451168 | NA | Used to align reads to human reference |
| software, algorithm | Picard v1.108 | Broad Institute; https://broadinstitute.github.io/picard/. |
NA | Identify duplicate reads |
| software, algorithm | The Genome Analysis Toolkit (GATK) v2.8.1 |
PMCID: PMC2928508 | NA | locally realign reads to known indels and recalibrate base quality scores |
| software, algorithm | MuTect v1.1.4 | PMCID: PMC3833702 | NA | Identiy somatic point mutations between the tumour and matched normal |
| other | WES/WGS raw sequencing data |
This paper | NCBI GEO Accession: PRJNA327240 |
Raw FASTQ files for WES and WGS performed in this study |