Rasal2^{Tn(pb-Act-RFP)1Xwu}

A piggyBac (PB) transposon element derived from cabbage looper moth Trichoplusia ni is introduced to intron 3 of the gene. Its components were introduced by coinjection of mouse oocytes with circular piggyBac donor and helper plasmids. The donor plasmid contained the piggyBac transposon element in which the transposase sequence was replaced by a beta actin (Actb) promoter-driven red flourescent protein (RFP) reporter gene flanked by piggyBac-specific inverted terminal repeat sequences. The helper plasmid employs the Actb promoter to drive expression of a piggyBac transposase gene lacking the terminal inverted repeat sequences required for its transposition. The insertion contains the RFP expression unit flanked by transposon (donor) and transposase (helper) sequences. RFP is flanked by transposon donor and transposase helper sequences in the final construct. Expression of Rasal2 is decreased by more than 90% in some tissues. (J:101977)