Direct modification of spermatogonial stem cells using lentivirus vectors in vivo leads to efficient generation of transgenic rats

Spermatogonial stem cells(SSCs)transmit genetic information to the next progeny in males.Thus,SSCs are a potential target for germ I i ne modifications to gen erate tran sgenic an imals.In this study,we report a technique for the gen erati on of tran sgenic rats by in vivo manipulation of SSCs with a high success rate.SSCs in juvenile rats were transduced in vivo with high titers of lentivirus harbori ng enhan ced green fluoresce nt protei n and mated with wild-type females to create foun der rats.These founder rats expressed the transgene and passed on the transgene with an overall success rate of 50.0%.Subsequent generations of progeny from the founder rats both expressed and passed on the transgene.Thus,direct modification of SSCs in juvenile rats is an effective means of generating transgenic rats through the male germline.This technology could be adapted to larger animals,in which existing methods for gene modificatio n are in adequate or in applicable,resulti ng in the gen eration of tran sge nic an imals in a variety of species.