We generated ROSA26-targetted ES cells and demonstrated that a transgene can be inserted into the ROSA26 locus at high frequency. We injected the ROSA26-targeted ES cells into mouse blastocysts and obtained chimeric pups. We confirmed the germline transmission of a targeted ROSA26 allele. However, due to problems in our mouse facility, we lost the mouse line. We are currently repeating the blastocyst injection and trying to re-establish this mouse line. We will then establish the homozygous mouse line and will then examine the frequency of transgene integration by injecting Cre and an exchange vector into fertilized eggs. We can then test whether the transgene can be induced in these embryos and adult mice in the absence of tetracycline.
| Nishiyama, Akira; Xin, Li; Sharov, Alexei A et al. (2009) Uncovering early response of gene regulatory networks in ESCs by systematic induction of transcription factors. Cell Stem Cell 5:420-33 |
