OGG1 Knockout HAP1 Cell Line

The choice depends on whether you are studying OGG1 (8-oxoguanine DNA glycosylase)'s role as the principal repair enzyme for oxidative DNA damage or its functions in oxidative stress response. The Knockout line is the standard tool for asking whether OGG1 is required for removing 8-oxoG lesions — OGG1 is a base excision repair glycosylase that recognizes and excises 8-oxoguanine paired with cytosine, preventing G→T transversions characteristic of oxidative mutagenesis. Overexpression is useful for studying OGG1 in oxidative stress protection. For DNA damage research, the EDITGENE OGG1 Knockout in HAP1 enables study of oxidative DNA damage repair. MTH1 (NUDT1, sanitizing 8-oxo-dGTP) and MUTYH (removing adenine from A:8-oxoG) provide partial functional overlap with OGG1. Rescue with wild-type or catalytically-dead OGG1 is the standard specificity control. The knockout is valuable for testing OGG1 inhibitors (TH5487, SU0268) being explored as anti-inflammatory and emerging cancer therapeutics.

Primary applications: • 8-oxoG repair: 8-oxoG accumulation analysis by immunofluorescence (anti-8-oxoG antibody) or LC-MS quantification. • Mutation rate analysis: G→T transversion mutational signatures in whole-exome sequencing. • Oxidative stress response: H2O2, paraquat, or KBrO3-induced cellular toxicity given OGG1's role in oxidative damage repair. • OGG1 inhibitor specificity: critical genetic control for TH5487, SU0268, and other OGG1-targeting compounds in inflammation and cancer drug development. EDITGENE recommends this model for researchers investigating oxidative DNA damage repair and OGG1-targeted therapeutic mechanisms.

Yes. OGG1 rescue experiments are well-established for BER research: • Construct design: use a codon-modified OGG1 sequence with a small C-terminal tag (FLAG, HA). OGG1 has glycosylase domain and DNA-binding helix-hairpin-helix motifs — preserve all elements. • Catalytically-dead rescue: K249Q (catalytic lysine) mutation abolishes 8-oxoG excision activity and is the standard specificity control. • AP lyase-deficient rescue: separate mutations affect AP lyase versus glycosylase activities — enabling dissection of dual functions. • Functional readout: rescue should restore 8-oxoG removal (LC-MS or anti-8-oxoG IF) and oxidative damage tolerance. HAP1-specific considerations: • Diploidization: HAP1 cells gradually diploidize during extended culture — confirm ploidy by flow cytometry at the time of phenotypic assay. • Integration site sensitivity: position effects on transgene expression are more pronounced in near-haploid backgrounds; generating multiple independent rescue clones is strongly recommended. • Transduction efficiency: HAP1 transduces with lentivirus at moderate efficiency — increase MOI compared to standard immortalized lines.