Use of tRNA gene barriers improves stability of transgene expression in CHO cells

Instability of transgene expression is a major challenge for the biopharmaceutical industry, which can impact yields and regulatory approval. Some tRNA genes (tDNAs) can resist epigenetic silencing, the principal mechanism of expression instability, and protect adjacent genes against the spread of repressive heterochromatin. We have taken two naturally occurring clusters of human tDNAs and tested their ability to reduce epigenetic silencing of transgenes integrated into the genome of Chinese hamster ovary (CHO) cells. We find sustained improvements in productivity both in adherent CHO‐K1 cells and in an industrially relevant CHO‐DG44 expression system (Apollo X, FUJIFILM Diosynth Biotechnologies). We conclude that specific tDNA clusters offer potential to mitigate the widespread problem of production instability. Graphical and Lay Summary A major challenge for the biopharmaceutical industry is instability of transgene expression, where epigenetic repression silences genes encoding high‐value products. We demonstrate that specific tRNA genes (tDNAs) can mitigate this widespread problem in Chinese hamster ovary (CHO) cells.