DTWD1 Knockout Cell Line (HEK293)

Important nomenclature clarification: DTWD1 (DTW domain containing 1) was historically annotated through homology, but biochemical characterization (Takakura et al. Nature Communications 2019) has established that DTWD1 is a tRNA aminocarboxypropyltransferase that catalyzes acp³U (3-(3-amino-3-carboxypropyl)uridine) modification at tRNA position 20 — DTWD1 is NOT a pseudouridine synthase despite some annotation databases suggesting this. The choice between knockout and overexpression depends on whether you are studying acp³U tRNA modification biology in human cells. The Knockout line is the standard tool for asking whether DTWD1 is required for acp³U modification at tRNA position 20 — DTWD1 uses S-adenosylmethionine (SAM) as the aminocarboxypropyl donor; DTWD1 and DTWD2 are the human homologs of E. coli TapT. For tRNA modification research, the EDITGENE DTWD1 Knockout in HEK293 enables study of acp³U modification biology. This product complements the parallel DTWD1 Knockout in AML12 (mouse hepatocyte, also available) and the DTWD1 & DTWD2 Double Knockout in HEK293 (also available) for cross-species and paralog-specific studies. Rescue with wild-type or catalytically-dead DTWD1 enables structure-function studies. The knockout is valuable for studying tRNA modification, translation fidelity, and emerging tRNA modification-related cancer biology — DTWD1 dysregulation has been reported in several malignancies.

Primary applications: • acp³U tRNA modification: cellular tRNA acp³U levels by HPLC-MS analysis in DTWD1-null versus rescued cells. • Translation fidelity: ribosome profiling and translation accuracy analysis given acp³U's role in tRNA function. • Heat stress response: cellular growth and tRNA stability under heat stress given acp³U's role in tRNA thermal stabilization. • Cross-species comparison: parallel analysis with DTWD1 Knockout in AML12 (mouse, also available) for cross-species studies. EDITGENE recommends this HEK293-based model for biochemical DTWD1 research; the parallel double knockout enables systematic acp³U pathway dissection.

Yes. DTWD1 rescue experiments require attention to acp³U transferase architecture: • Construct design: use a codon-modified DTWD1 sequence with a small C-terminal tag (FLAG, HA). DTWD1 has the DTW domain containing the active site — preserve domain integrity. • Catalytically-dead rescue: active site residue mutations abolish aminocarboxypropyltransferase activity and serve as the standard specificity control. • Functional readout: rescue should restore tRNA acp³U levels at position 20 measured by HPLC-MS. HEK293 transduces efficiently with lentivirus and supports stable rescue line generation.