Single copy shRNA configuration for ubiquitous gene knockdown in mice.
RNA interference through the expression of small hairpin RNA (shRNA) molecules has become a very promising tool in reverse mouse genetics as it may allow inexpensive and rapid gene function analysis in vivo. However, the prerequisites for ubiquitous and reproducible shRNA expression are not well defined. Here we show that a single copy shRNA-transgene can mediate body-wide gene silencing in mice when inserted in a defined locus of the genome. The most commonly used promoters for shRNA expression, H1 and U6, showed a comparably broad activity in this configuration. Taken together, the results define a novel approach for efficient interference with expression of defined genes in vivo. Moreover, we provide a rapid strategy for the production of gene knockdown mice combining recombinase mediated cassette exchange and tetraploid blastocyst complementation approaches.
Seibler
Jost
J
Artemis Pharmaceuticals GmbH Neurather Ring 1, 51063 Cologne, Germany. j.seibler@artemispharma.de
Küter-Luks
Birgit
B
Kern
Heidrun
H
Streu
Sandra
S
Plum
Leona
L
Mauer
Jan
J
Kühn
Ralf
R
Brüning
Jens C
JC
Schwenk
Frieder
F
D000818:Animals; D002478:Cells, Cultured; D018628:Gene Dosage; D006801:Humans; D049409:Luciferases, Firefly / Q000032:analysis / Q000235:genetics; D051379:Mice / Q000235:genetics*; D016254:Mutagenesis, Insertional; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D022661:RNA, Untranslated; D011956:Receptors, Cell Surface / Q000235:genetics; D054411:Receptors, Leptin; D045522:Recombinases / Q000378:metabolism; D019076:Transgenes; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D011506:Proteins; D034741:RNA, Small Interfering; D022661:RNA, Untranslated; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin; D045522:Recombinases; D049409:Luciferases, Firefly; D001616:beta-Galactosidase