DCAF16 Knockout Cell Line (Hela)

The choice depends on whether you are studying DCAF16 (DDB1- and CUL4-associated factor 16)'s role as a CRL4 substrate receptor or modeling its emerging applications in covalent molecular glue degraders. The Knockout line is the standard tool for asking whether DCAF16 is required for these processes — DCAF16 is one of the ~70 DCAF (DDB1- and CUL4-associated factor) substrate receptors that, together with DDB1 and CUL4A/4B, form CRL4 E3 ubiquitin ligase complexes; DCAF16 contains a key surface cysteine (C58, C107, or other) recently characterized as a target for covalent molecular glue degraders. Overexpression is useful for studying DCAF16 in heterologous expression contexts. For targeted protein degradation research, the EDITGENE DCAF16 Knockout in HeLa is uniquely valuable — DCAF16 has emerged as one of the most promising targets for covalent molecular glue degrader (CovGD) development, with recent studies (Henning et al. 2022; Pluciński et al. 2023) showing that small molecules can covalently engage DCAF16 cysteines to recruit neo-substrates for degradation. Rescue with wild-type or cysteine-mutant (C58A, C107A) DCAF16 enables systematic characterization of covalent molecular glue mechanism. The knockout is a critical specificity tool for emerging DCAF16-recruiting covalent degraders and broader exploration of cysteine-reactive molecular glue platforms — DCAF16 is at the forefront of the covalent TPD field.

Primary applications: • CRL4-DCAF16 substrate identification: ubiquitinome analysis in DCAF16-null versus rescued cells to identify endogenous DCAF16 substrates. • Covalent molecular glue specificity: critical genetic control for DCAF16-recruiting covalent molecular glue degraders — these compounds should have no degradation activity in DCAF16-null cells. • Cysteine reactivity studies: rescue with cysteine-to-alanine mutants (C58A, C107A) for systematic characterization of cysteine-targeting covalent molecular glues. • Targeted protein degradation platform: development of new DCAF16-recruiting molecular glue degraders and PROTACs. EDITGENE recommends this model as a critical specificity tool for the rapidly emerging field of covalent molecular glue degraders targeting DCAF16.

Yes, and rescue experiments are uniquely powerful for covalent molecular glue research: • Construct design: use a codon-modified DCAF16 sequence with a small C-terminal tag (FLAG, HA). DCAF16 contains the WD40-like substrate-receptor region with key surface cysteines (C58, C107, others) — preserve all elements. • Cysteine-to-alanine rescue: C58A, C107A, and other cysteine mutations enable systematic characterization of which cysteines are required for covalent molecular glue recruitment — this is the gold-standard approach for understanding covalent ligand-DCAF16 mechanism. • Functional readout: rescue should restore CRL4-DCAF16 complex assembly and substrate ubiquitination; covalent molecular glue degrader activity requires the specific cysteine being engaged. HeLa transduces efficiently with lentivirus and supports systematic cysteine mutation analysis for the rapidly emerging covalent molecular glue degrader field.