For the Stk4tm1.1Rjo (mst1) targeted mutation, a targeting vector containing an FRT flanked NEO cassette was utilized in the construction of this mutant. This selection cassette was inserted downstream of exon 5 and loxP sites were inserted flanking exons 4 and 5. The construct was electroporated into (C57BL/6Brd-Tyrc-Brd x 129S6/SvEvTac)F1-derived RJ2.2 embryonic stem (ES) cells. ES cells containing the construct were aggregated with CD1 morulae and the resulting chimeric males were bred with CD1 females for germline transmission. The mice were then bred to a FLPe recombinase expressing strain to remove the selection cassette. Heterozygotes that retained the loxP site flanked exons 4 and 5 were then intercrossed to generation homozygotes.
For the Stk3tm1.1Rjo (mst2) targeted mutation, a targeting vector containing an FRT flanked NEO cassette was utilized in the construction of this mutant. This selection cassette was inserted downstream of exon 6 and loxP sites were inserted flanking exons 5 and 6. The construct was electroporated into (C57BL/6Brd-Tyrc-Brd x 129S6/SvEvTac)F1-derived RJ2.2 embryonic stem (ES) cells. ES cells containing the construct were aggregated with CD1 morulae and the resulting chimeric males were bred with CD1 females for germline transmission. The mice were then bred to a FLPe recombinase expressing strain to remove the selection cassette. Heterozygotes that retained the loxP site flanked exons 5 and 6 were then intercrossed to generation homozygotes.
The single mutant lines were then intercrossed to obtain this double mutant strain, which was maintained on a mixed background consisting of CD1, C57BL/6 and 129.
Upon arrival at The Jackson Laboratory, the mice were crossed to C57BL/6J (Stock No. 000664) at least once to establish the colony.
