Effect of Dox-induced Cas13 expression in stable cell lines on the mouse transcriptome and co-cultured human PSCs

In the mouse EpiSCs-iCas13 stable cell line, addition of doxycycline induces Cas13 expression, which further triggers collateral effects and slows down cell proliferation. In co-culture of mouse and human cells, under normal conditions, mouse EpiSCs hold a proliferative advantage, whereas human PSCs are at a disadvantage and show almost no growth. Upon doxycycline treatment in the co-culture, Cas13 expression in mouse cells triggers collateral effects that reduce their proliferation, thereby providing a relative advantage for human PSCs. As a result, the proliferation of human PSCs is markedly enhanced, showing a growth pattern more comparable to that observed in separate cultures of human PSCs. Overall design: Cells from separate cultures and co-cultures, with or without doxycycline treatment, were collected on day 4. Total RNA was extracted, followed by RNA-seq and subsequent transcriptome analysis.