ATG12 Knockout HAP1 Cell Line

The choice depends on whether you are studying ATG12's role as a ubiquitin-like protein in the ATG12-ATG5 conjugation system for autophagy or modeling autophagy deficiency. The Knockout line is the standard tool for asking whether ATG12 is required for canonical autophagy — ATG12 is a ubiquitin-like protein that is conjugated to ATG5 via ATG7 (E1)-ATG10 (E2) cascade, with the ATG12-ATG5 conjugate functioning as the E3-like ligase for LC3 lipidation; ATG12 is essential for canonical autophagy. Overexpression is useful for studying ATG12 gain-of-function effects. For autophagy research, the EDITGENE ATG12 Knockout in HAP1 is the gold-standard autophagy-deficient haploid cell model — ATG12 KO abolishes LC3-II formation and canonical macroautophagy. This product complements the parallel ATG7 Knockout in HEK293 (also available) for systematic autophagy machinery dissection. Rescue with wild-type or conjugation-deficient ATG12 (G140A C-terminal glycine for ATG5 conjugation) enables structure-function studies. The knockout is valuable for studying autophagy biology and emerging ATG12-related apoptosis regulation (an unusual autophagy-independent role of ATG12).

Primary applications: • Autophagy assays: LC3-I/LC3-II conversion and p62 accumulation analysis in ATG12-null cells. • ATG12-ATG5 conjugation system: ATG5 conjugation analysis given ATG12 KO abolishes ATG12-ATG5 conjugate formation. • Cross-background validation: parallel analysis with ATG7 KO in HEK293 (also available). • Autophagy-independent ATG12 functions: emerging non-autophagy roles of ATG12 in apoptosis regulation. EDITGENE recommends this HAP1-based model for systematic autophagy machinery dissection complementary to the ATG7/HEK293 product.

Yes. ATG12 rescue experiments are gold-standard for autophagy research: • Construct design: use a codon-modified ATG12 sequence with a small C-terminal tag — but note that ATG12 is conjugated to ATG5 via its C-terminal glycine (G140); C-terminal tags may interfere with conjugation. N-terminal tag is preferred. • Conjugation-deficient rescue: G140A mutation abolishes ATG5 conjugation — gold-standard specificity control. • Functional readout: rescue should restore ATG12-ATG5 conjugation and LC3-II formation. 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.