RHBDF1 Knockout Cell Line (HEK293)

The choice depends on whether you are studying RHBDF1 (iRhom1)'s role as a broadly expressed regulator of ADAM17/TACE-mediated ectodomain shedding — distinct from iRhom2's more tissue-restricted expression — or its functions in keratinocyte biology and cancer. The Knockout line is the standard tool for asking whether iRhom1 is required for ADAM17 maturation in non-immune cell contexts where it is the dominant iRhom paralog. Overexpression is useful for studying iRhom1's tissue-specific TACE regulation. For ADAM17 regulation research, the EDITGENE RHBDF1 Knockout in HEK293 is a mechanistic platform — iRhom1 is the principal iRhom in many epithelial contexts including HEK293. This product complements the parallel RHBDF1 Knockout in A-549 and the RHBDF1 & RHBDF2 Double Knockouts; HEK293 is preferred for biochemistry and structure-function studies. iRhom1 is essential for ADAM17 ER exit and Golgi maturation — single iRhom1 knockout in HEK293 substantially reduces functional ADAM17. Rescue with wild-type or non-functional iRhom1 variants enables structure-function studies.

Primary applications: • ADAM17 ER exit: pro-ADAM17 retention versus surface ADAM17 analysis to assess iRhom1's contribution to TACE trafficking. • Combined iRhom analysis: combination with RHBDF2 knockdown or knockout (or use of the double knockout) to dissect iRhom1-specific versus pan-iRhom functions. • ADAM17 substrate panel: ectodomain shedding analysis for the broader ADAM17 substrate repertoire (>80 substrates including IL-6R, L-selectin, others). • Structure-function: iRhom1 variant rescue with the high-transfection HEK293 background. EDITGENE recommends this HEK293-based model for iRhom1 biochemistry and ADAM17 trafficking research; the parallel A-549 knockout and double knockouts complement for paralog and cancer studies.

Yes. iRhom1 rescue experiments are well-established for ADAM17 trafficking research: • Construct design: use a codon-modified RHBDF1 sequence with a small C-terminal tag (FLAG, HA). iRhom1 has the rhomboid family 7-transmembrane architecture, lacking catalytic activity as a pseudoenzyme. • N-terminal domain mutant rescue: phospho-site mutations enable studies of iRhom1 regulation distinct from iRhom2. • Paralog rescue with iRhom2: rescue interpretation considers RHBDF2 expression — in single iRhom1 KO, residual iRhom2 may partially support ADAM17. • Functional readout: rescue should restore ADAM17 ER exit and Golgi maturation. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.