| First Author | Sharma N | Year | 2008 |
| Journal | Nucleic Acids Res | Volume | 36 |
| Issue | 11 | Pages | e67 |
| PubMed ID | 18499713 | Mgi Jnum | J:173065 |
| Mgi Id | MGI:5009526 | Doi | 10.1093/nar/gkn298 |
| Citation | Sharma N, et al. (2008) Regulated gene insertion by steroid-induced PhiC31 integrase. Nucleic Acids Res 36(11):e67 |
| abstractText | Nonviral integration systems are widely used genetic tools in transgenesis and play increasingly important roles in strategies for therapeutic gene transfer. Methods to efficiently regulate the activity of transposases and site-specific recombinases have important implications for their spatiotemporal regulation in live transgenic animals as well as for studies of their applicability as safe vectors for genetic therapy. In this report, strategies for posttranslational induction of a variety of gene-inserting proteins are investigated. An engineered hormone-binding domain, derived from the human progesterone receptor, hPR891, and specifically recognized by the synthetic steroid mifepristone, is fused to the Sleeping Beauty, Frog Prince, piggyBac and Tol2 transposases as well as to the Flp and PhiC31 recombinases. By analyzing mifepristone-directed inducibility of gene insertion in cultured human cells, efficient posttranslational regulation of the Flp recombinase and the PhiC31 integrase is documented. In addition, fusion of the PhiC31 integrase with the ER(T2) modified estrogen receptor hormone-binding domain results in a protein, which is inducible by a factor of 22-fold and retains 75% of the activity of the wild-type protein. These inducible PhiC31 integrase systems are important new tools in transgenesis and in safety studies of the PhiC31 integrase for gene therapy applications. |
