DTWD1 & DTWD2 Knockout HEK293 Cell Line

The choice depends on whether you are studying combined DTWD1/DTWD2 function or distinguishing complete acp³U-deficient phenotypes. The Double Knockout line is uniquely valuable for asking whether DTWD1/DTWD2 are required for global acp³U tRNA modification — DTWD1 and DTWD2 catalyze acp³U at different tRNA positions (DTWD1 at position 20, DTWD2 at position 20a); combined loss eliminates acp³U at both positions, generating completely acp³U-deficient tRNAs. Single-isoform rescue (DTWD1 alone or DTWD2 alone) in the double knockout enables position-specific functional dissection. For tRNA modification research, the EDITGENE DTWD1 & DTWD2 Double Knockout in HEK293 is the gold-standard genetic tool — Takakura et al. (Nature Communications 2019) reported that DTWD1+DTWD2 double knockout cells exhibit growth retardation, indicating that acp³U is physiologically important in mammals. Single-isoform rescue is the gold-standard experimental design for paralog dissection. The double knockout is uniquely valuable for studying acp³U-dependent tRNA biology, translation fidelity, and emerging tRNA modification-related disease research.

Primary applications: • Complete acp³U elimination: global acp³U tRNA modification analysis — double KO eliminates DTWD1- and DTWD2-dependent modifications at tRNA positions 20 and 20a. • Single-isoform rescue: re-introduction of DTWD1 alone or DTWD2 alone enables position-specific functional dissection — gold-standard experimental design. • Growth retardation analysis: characterization of acp³U-deficient cellular phenotypes given the reported growth retardation in DTWD1+DTWD2 double KO cells. • Translation analysis: ribosome profiling and translation fidelity in completely acp³U-deficient tRNA context. EDITGENE recommends this double knockout as the gold-standard genetic tool for acp³U-targeted tRNA modification research.

Yes, and rescue experiments are uniquely powerful in this double knockout: • Single-isoform rescue: re-introduction of DTWD1 alone or DTWD2 alone in the double knockout enables position-specific functional dissection (DTWD1 modifies position 20; DTWD2 modifies position 20a) — gold-standard experimental design for paralog dissection. • Construct design: use codon-modified DTWD1 or DTWD2 sequences with small C-terminal tags (FLAG, HA). Preserve DTW domain integrity in each paralog. • Catalytically-dead rescue: active site mutations enable separation of catalytic from structural functions. • Functional readout: rescue should restore position-specific acp³U levels and reverse the growth retardation phenotype reported in double knockout cells. HEK293 transduces efficiently with lentivirus and supports systematic isoform-specific rescue experiments.