SQSTM1 Knockout Cell Line (Hela)

The choice depends on whether you are studying p62's role as a selective autophagy receptor or its functions in NF-κB and Nrf2/Keap1 signaling. The Knockout line is the standard tool for asking whether p62 is required for selective autophagy of ubiquitinated cargo (aggrephagy, mitophagy, xenophagy). Overexpression is useful for studying p62's role in inclusion body formation or for testing p62 condensation/LLPS behavior. For autophagy research, the EDITGENE p62 Knockout in HeLa is a workhorse tool — HeLa has been the principal cell line for selective autophagy mechanism studies, with extensive published p62-dependent phenotypes. This product complements the parallel p62 Knockout in HEK293; HeLa is preferred for imaging-based aggrephagy studies given its flat morphology and well-characterized autophagy machinery. Rescue with wild-type, UBA-mutant (ubiquitin-binding-deficient), or LIR-mutant (LC3-binding-deficient) p62 enables comprehensive selective autophagy dissection.

Primary applications: • Selective autophagy assays: aggrephagy (puromycin-induced aggregates), mitophagy (CCCP-induced), and xenophagy assays in the absence of p62. • Imaging-based autophagy: LC3 puncta, p62 puncta dynamics, and autophagic flux measurement using imaging — HeLa's flat morphology supports high-resolution imaging. • Inclusion body formation: protein aggregate accumulation assays following proteasome inhibition. • Cargo recognition studies: rescue with UBA-mutant or LIR-mutant p62 to dissect ubiquitin-binding versus LC3-engagement functions. EDITGENE recommends this model for researchers investigating selective autophagy, particularly imaging-based studies of p62-dependent cargo sorting.

Yes. p62 rescue experiments in HeLa are well-established for autophagy research: • Construct design: use a codon-modified SQSTM1 sequence with a small N- or C-terminal tag (FLAG, HA, GFP for imaging). p62 contains multiple functional domains (PB1, ZZ, TB, LIR, KIR, UBA) — small tags should be used to minimize domain disruption. • Domain-specific rescue panel: UBA mutation (K7A or domain deletion) abolishes ubiquitin binding; LIR mutation (W340A) abolishes LC3 binding — these constitute the standard selective autophagy specificity controls. • PB1-mutant rescue: PB1 domain mutations (D69A) abolish p62 oligomerization, separating LLPS-dependent functions from non-condensate functions. • Functional readout: rescue should restore selective autophagy of aggregated substrates and p62 puncta dynamics imaged in HeLa's flat morphology background. HeLa transduces efficiently with lentivirus; its imaging compatibility makes it particularly well-suited for fluorescent p62 rescue construct studies.