High-efficiency gene knockdown using chimeric ribozymes in fish embryos.
We report an effective gene knockdown technique in rainbow trout embryos using additional RNA components combined with ribozymes (R(z)s). Chimeric R(z)s (tR(z)Cs) containing tRNA(Val), R(z) against GFP, and a constitutive transport element were microinjected into transgenic embryos. tR(z)Cs induced greater gene interference than R(z)s alone. Control tR(z)Cs did not affect unpaired bases of target RNA, and the tR(z)C did not interfere with non-relevant gene expression, suggesting that the tR(z)C-mediated gene-interference effects were sequence-specific. Furthermore, the tR(z)C-containing expression vector specifically suppressed target GFP expression in transgenic trout. tR(z)Cs enhance R(z) cleavage and could therefore be powerful tools for studying unknown gene function in vertebrates.
Boonanuntanasarn
Surintorn
S
School of Animal Production Technology, Institute of Agricultural Technology, Suranaree University of Technology, 111 University Avenue, Muang, Nakhon Ratchasima 30000, Thailand.
Takeuchi
Toshio
T
Yoshizaki
Goro
G
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology* / Q000235:genetics; D002678:Chimera / Q000196:embryology / Q000235:genetics; D005399:Fishes / Q000196:embryology* / Q000235:genetics*; D020868:Gene Silencing; D005818:Genetic Engineering / Q000379:methods*; D016337:RNA, Catalytic / Q000235:genetics*
D016337:RNA, Catalytic