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To allow for incorporation of ONBY in eukaryotic systems, an Mb PylRS was engineered for ONBY incorporation through three rounds of alternating positive and negative selection using a library of mutants with L270, Y271, L274, N311 and N313 mutated to all possible amino acids
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introduction of the anti-ketoglutarate lyase gene into the TME, which neutralizes toxic metabolites like Kyn and breaks down inhibitory metabolites
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