Gt(ROSA)26Sor^{tm6(RNAi:Gsk3a)Rkuhn}

Recombinase mediated cassette exchange (RMCE) was used to generate an allele where upon expression of Cre recombinase a knockdown RNAi is generated to Gsk3a mRNA. Short hairpin RNA was designed with a loxP-flanked stop cassette in the loop portion of the RNAi between the sense and anti-sense sequences. The RNAi was put under the control of a human U6 promoter and the cassette was placed downstream of a splice acceptor and a frt-flanked neomycin resistance gene. RMCE was then performed on ES cells containing the Gt(ROSA)26tm1Rkuhn allele. Upon expression of cre recombinase, protein expression of Gsk3a was reduced by 84%. (J:133340)