SIRT7 Knockout HAP1 Cell Line

The choice depends on whether you are studying SIRT7's role as a nucleolar NAD⁺-dependent deacetylase regulating ribosomal RNA transcription or its emerging functions in DNA damage response, metabolism, and aging biology. The Knockout line is the standard tool for asking whether SIRT7 is required for H3K18ac deacetylation, RNA polymerase I transcription regulation, or its other reported functions. Overexpression is useful for studying SIRT7 in cancer contexts where it has been reported to be upregulated. For sirtuin research, the EDITGENE SIRT7 Knockout in HAP1 provides a clean genetic background for dissecting SIRT7-specific functions among the seven mammalian sirtuins. Rescue with wild-type or catalytically-dead (H187Y) SIRT7 is the standard specificity control. SIRT7 has emerging interest in aging, NAD⁺ biology, and cancer — the knockout serves as a critical specificity tool for SIRT7-selective compounds in development.

Primary applications: • H3K18ac deacetylation: histone H3K18 acetylation Western blot/ChIP analysis at SIRT7 target gene promoters. • Ribosomal RNA transcription: 47S pre-rRNA quantification by qPCR to assess SIRT7's effect on Pol I transcription. • NAD⁺-dependent activity: in vitro deacetylation assays with NAD⁺ supplementation and varied conditions. • Cancer biology: proliferation, transformation, and stress response phenotypes given SIRT7's reported tumor-supportive functions. EDITGENE recommends this model for researchers investigating sirtuin biology, nucleolar function, NAD⁺-dependent deacetylase mechanisms, and SIRT7-targeted compound development.

Yes. SIRT7 rescue experiments are well-established for sirtuin research: • Construct design: use a codon-modified SIRT7 sequence with a small C-terminal tag (FLAG, HA). SIRT7 has a central catalytic domain with N- and C-terminal extensions critical for nucleolar localization and substrate recognition. • Catalytically-dead rescue: the H187Y mutation abolishes NAD⁺-dependent deacetylase activity and is the standard specificity control. • Nucleolar localization validation: confirm nucleolar localization by fibrillarin co-staining before functional assays. • Functional readout: rescue should restore H3K18ac levels at SIRT7 target promoters and ribosomal RNA transcription. 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.