[Intercellular mRNA transfer investigated by a Cre-loxP system].

Objective To study the process and significance of intercellular mRNA transfer by using a Cre-loxP system. Methods Cre-expressing donor cells and loxP-DsRed-loxP-eGFP-expressing recipient cells were established. Co-culture and TranswellTM co-culture of both donor and recipient cells were analyzed with a separate culture of recipient cells only as controls. After 72 hours, the eGFP+ recipient cells in each group were observed and analyzed. Immunofluorescence staining was used to detect Cre expression in eGFP+ cells. The effects of donor cell death, cell membrane nanotubes, and ROCK signaling pathway on Cre mRNA transfer were explored. Results After co-culture for 72 hours, the eGFP+ cells were only observed in co-culture group, but not in the recipient cells alone group and the TranswellTM co-culture group, indicating that cell-cell contact could mediate Cre mRNA transfer. Immunofluorescence staining showed that Cre mRNA could be translated into proteins and transported to the nucleus in eGFP+ recipient cells. Moreover, the transport of Cre mRNA between living cells could be partially blocked by membrane nanotubes or ROCK signaling pathway inhibitors. Conclusion The mRNA can be exchanged among living cells through intercellular contact, thus leading to the changes of biological functions in the recipient cells.