Retrovirus-mediated mRNA transfer (RMT) combines the advantageous features of retrovirus-mediated cell targeting and entry with the controlled transfer of mRNAs. We have recently exploited this strategy for the dose-controlled transfer of recombinases and DNA transposases, avoiding cytotoxicity and potential insertional mutagenesis. Further applications can be envisaged, especially when low expression levels are sufficient to modify cell fate or function. Here we describe a step-by-step protocol for the generation of RMT vector particles, their titration and their application in a model cell line.