Mass spectrometry confirmed Ala mis-incorporation in polyQ, demonstrating misincorporation at each position in polyQ at a level of ~10% Ala at Gln codons. Cells expressing the missense suppressor tRNAs showed no growth defect or significant changes in cytotoxicity. Our findings demonstrate that tRNA-dependent missense suppression of glutamine codons is well-tolerated in cells has the potential to modify and significantly reduce protein aggregation that causes HD.