Heritable and stable gene knockdown in rats.
The rat has served as an excellent model for studies on animal physiology and as a model for human diseases such as diabetes and alcoholism; however, genetic studies have been limited because of the inability to knock out genes. Our goal was to produce heritable deficiencies in specific gene function in the rat using RNA interference to knock down gene expression in vivo. Lentiviral-mediated transgenesis was used to produce rats expressing a short hairpin RNA targeting Dazl, a gene expressed in germ cells and required for fertility in mice. Germ-line transmission of the transgene occurred, and its expression correlated with significant reductions in DAZL protein levels and male sterility, and the knockdown was stable over multiple generations (F(1)-F(3)). This study demonstrates an efficient system by which directed reverse genetic analysis can now be performed in the rat.
Dann
Christina Tenenhaus
CT
The Cecil H. and Ida Green Center for Reproductive Biology Sciences, Department of Pharmacology, University of Texas Southwestern Medical Center, 6001 Forest Park Road, Dallas, TX 75390-9051, USA. christina.dann@utsouthwestern.edu
Alvarado
Alma L
AL
Hammer
Robert E
RE
Garbers
David L
DL
D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D005260:Female; D005821:Genetic Techniques; D016086:Lentivirus / Q000235:genetics; D008297:Male; D008957:Models, Genetic; D008969:Molecular Sequence Data; D034622:RNA Interference; D016601:RNA-Binding Proteins / Q000235:genetics / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D019076:Transgenes
C512808:DAZL protein, rat; D016601:RNA-Binding Proteins