| First Author | Valencik ML | Year | 2001 |
| Journal | Transgenic Res | Volume | 10 |
| Issue | 3 | Pages | 269-75 |
| PubMed ID | 11437283 | Mgi Jnum | J:70619 |
| Mgi Id | MGI:2137847 | Doi | 10.1023/a:1016601928465 |
| Citation | Valencik ML, et al. (2001) Codon optimization markedly improves doxycycline regulated gene expression in the mouse heart. Transgenic Res 10(3):269-75 |
| abstractText | Tetracycline regulated gene expression in transgenic animals is potentially a very powerful technique (Furth et al., 1994; Gossen & Bujard 1992). We have utilized this system in an attempt to overcome the perinatal lethality resulting from constitutive transgenic expression in the heart (Valencik & McDonald, Am J Physiol Heart Circ Physiol 280: H361-H367). We found that compound hemizygous animals created by mating selected reverse tetracycline transactivator (rtTA) and transresponder (TR) lines display tightly regulated TR expression in the heart. However, we identified two fundamental problems. First, codon usage bias appeared to severely limit the expression of the rtTA driven by the cardiac alpha-myosin heavy chain promoter. Second, co-injection of rtTA and TR transgenes led to compound hemizygous animals that exhibited unregulated TR gene expression. Codon optimization of the rtTA construct leads to marked improvement (increasing the average induction from 20-fold to 832-fold) in cardiac myocyte expression. The resulting opt-rtTA lines can be bred to homozygosity, facilitating rapid screening of F0 TR animals for doxycycline regulated transgene expression. |
