Silencing efficiency differs among tissues and endogenous microRNA pathway is preserved in short hairpin RNA transgenic mice

• Published in: FEBS letters 2009-01, Vol.583 (1), p.213-218
• Main Authors: Sasaguri, Hiroki, Mitani, Tasuku, Anzai, Masayuki, Kubodera, Takayuki, Saito, Yuki, Yamada, Hiromi, Mizusawa, Hidehiro, Yokota, Takanori
• Format: Article
• Language: English
• Subjects: AAV; adeno-associated virus; Ago2; Animals; Argonaute-2; cDNA; Cerebrum - metabolism; complementary DNA; copper/zinc superoxide dismutase; double stranded RNA; dsRNA; embryonic stem; GAPDH; Gene Silencing; glyceraldehyde-3-phosphate dehydrogenase; Mice; Mice, Transgenic; MicroRNA; MicroRNAs - metabolism; miRNA; Nucleic Acid Conformation; Oversaturation; PBS; phosphate-buffered saline; Pol III; polymerase III; reverse transcription polymerase chain reaction; RISC; RNA interference; RNA-induced silencing complex; RNAi; RT-PCR; SDS; Short hairpin RNA; shRNA; Small interfering RNA; small nuclear RNA; snRNA; SOD1; sodium dodecyl sulfate; Superoxide Dismutase - genetics; Superoxide Dismutase-1; Transgenic mice
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2008.12.012

In short hairpin RNA (shRNA) transgenic mice, the tissue difference in gene silencing efficiency and oversaturation of microRNA (miRNA) pathway have not been well assessed. We studied these problems in our previously-reported anti-copper/zinc superoxide dismutase (SOD1) shRNA transgenic mice. Although there was a tissue difference (liver and skeletal muscle, >95%; central nervous system and lung, ∼80%), the target gene silencing was systemic and our anti-SOD1 shRNA transgenic mice recapitulated the SOD1-null mice. Neither endogenous miRNAs nor their target gene levels were altered, indicating the preservation of endogenous miRNA pathways. We think that the shRNA transgenic mice can be utilized for gene analysis.